Extraction - Science topic

Jhonny Vásquez Jiménez

Conventional chemical soil analysis can provide valuable insights into soil fertility by measuring key nutrient levels, pH, and other chemical properties. However, while it is a widely used method, its accuracy in assessing soil fertility can vary depending on several factors. Here are some considerations:

1. Nutrient Levels: Chemical soil analysis typically measures the concentrations of essential nutrients such as nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), sulfur (S), and micronutrients like iron (Fe), manganese (Mn), zinc (Zn), copper (Cu), boron (B), and molybdenum (Mo). While these measurements provide valuable information about soil nutrient status, they may not always reflect nutrient availability to plants. Soil nutrient availability can be influenced by factors such as soil pH, organic matter content, and soil texture, which may not be fully captured by chemical analysis alone.

2. pH and Soil Acidity/Alkalinity: Soil pH is a critical factor that influences nutrient availability, microbial activity, and plant growth. Conventional chemical analysis provides a reliable measurement of soil pH, but it may not account for localized variations within the soil profile or temporal changes over time. Additionally, soil pH alone may not fully predict nutrient availability, as other factors such as soil buffering capacity and root uptake mechanisms also play important roles.

3. Organic Matter Content: Chemical soil analysis typically does not directly measure soil organic matter content, which is a key determinant of soil fertility and productivity. While indirect methods such as loss on ignition (LOI) or Walkley-Black method can estimate organic matter content, they may not capture the full spectrum of organic matter types and their contributions to soil fertility.

4. Soil Biological Properties: Conventional chemical analysis focuses primarily on soil chemical properties and may not capture important biological indicators of soil fertility, such as microbial biomass, enzyme activity, and soil biodiversity. These biological properties are integral to nutrient cycling, organic matter decomposition, and soil health but are not typically assessed in routine chemical soil tests.

5. Interpretation and Management Recommendations: While chemical soil analysis provides quantitative data on soil nutrient levels and pH, interpreting these results and translating them into actionable management recommendations requires expertise and consideration of site-specific factors. Soil fertility management strategies should integrate chemical, physical, and biological aspects of soil health to optimize nutrient availability, crop productivity, and environmental sustainability.

In summary, while conventional chemical soil analysis is a valuable tool for assessing soil fertility, its accuracy and relevance depend on factors such as nutrient availability, soil pH, organic matter content, and biological properties. Integrating chemical analysis with other soil assessment methods, such as soil biological testing and field observations, can provide a more comprehensive understanding of soil fertility and inform effective soil management practices.

To proof that a small molecule is covalently bound to a polymer is very challenging. I was in similar situation when I tried to bind polyphenols covalently to chitosan. Generally, the polyphenols in your washing solution were either non-covalently bound or the covalent bond was cleaved during washing. The unremoved polyphenols are not necessarily bound covalently, as non-covalent interactions can still be too strong.

You could try to depolymerise your protein (e.g. by enzymatic hydrolysis) and then analyse the fragments (e.g. by LC-MS). If you find a fragment with the polyphenol bound covalently, you have a proof.

Ricarda Koopmann I'm quite satisfied with the purity and the quantity of my product in the organic phase. I believe it's fairly pure based on the IR spectra. Thank you very much for your response and advice

Lack of fit F test is used to test the hypothesis of model correctness: that is whether there is statistically significant model misspecification in not.

In RSM, the Lack of fit test is used to determine whether there is significant curvature in the experimental data: the first order model with interaction term is not able to capture adequate data variation and hence a second order model based design (CCD or Box Behnken type) is needed to fit the data coming from this particular region of the experimental space.

Details about the RSM process and statistical tests associated with it can be found in the following books:

1) Chemometrics: Experimental Design by Ed Morgan

2) Response Surface Methodology by Myers, Montogomery and Cook

Use Qubit or Bioanalyzer (or quantitative gel electrophoresis if nothing else) for your measurements if you can, phenol residues can give false positives with Nanodrop.

Essential oils are called so because they were originally thought to represent the very essence of the odor and flavor of the plant from which they are derived. The term "essential" in essential oils refers to the essence or characteristic fragrance of the plant that is captured in the concentrated hydrophobic liquid, which contains volatile chemical compounds extracted from plants.

When it comes to the type of GC/MS to use for trans-fatty acid analysis, I recommend a GC-MS system that offers high sensitivity, resolution, and accuracy. Look for a GC-MS instrument with a reliable chromatographic column and mass spectrometer detector to ensure precise identification and quantification of trans fatty acids in your food samples. Additionally, consider the software capabilities for data analysis and interpretation to enhance the efficiency of your analysis process.

Polysaccharides can be a common contaminant in bacterial DNA extraction, particularly when working with certain types of bacterial cells or tissues. These polysaccharides can interfere with downstream applications such as PCR or sequencing.

Lysozyme is sometimes used in DNA extraction protocols, particularly when dealing with bacterial cells or certain types of tissues. The amount of lysozyme required can vary depending on factors such as the type and quantity of starting material, the specific DNA extraction protocol being used, and the desired yield and purity of the extracted DNA.

Typically, lysozyme is added to a DNA extraction buffer or solution at a concentration ranging from 1 to 10 mg/ml. However, it's important to consult the specific protocol you're following, as the recommended concentration may vary.

For example, in some protocols for bacterial DNA extraction, a concentration of 1 mg/ml of lysozyme might be sufficient. In other cases, particularly when dealing with tougher cell walls or larger quantities of starting material, a higher concentration of lysozyme may be necessary.

It's also worth noting that lysozyme is often used in combination with other enzymes and reagents (such as proteinase K, SDS, or EDTA) to efficiently lyse cells and degrade proteins, releasing the DNA for subsequent purification steps.

hi Jonathan, thank you for your all response.

i am not referring to the paper exactly you mentioned, But of course I wanted to identify a Candidatus/uncultured bacteria. will go ahead with the 16S PCR and sequence the product....thank you again ...regards

No, you have to apply the Beer–Lambert law. (https://en.wikipedia.org/wiki/Beer%E2%80%93Lambert_law) However, this is only possible in a certain absorbance range (0.2 - 1.2, I think). Also, extinction coefficients found in literature are not sufficiently relieable. You have to determine it yourself by a calibration.

it may be possible to analyze cyclohexanone in C18-HPLC without extraction by optimizing the separation conditions, such as the flow rate, mobile phase composition, and column temperature. However, the selection of the appropriate column and mobile phase is crucial for the successful separation of cyclohexanone from other compounds in the matrix.

Hello,

I agree with Dr. Paul above, this happens due to formation of heteroduplexes. Your original band contains more than one product, with no noticeable difference in mobility, but the slow moving band is a heteroduplex. On cutting the expected band and Re-PCR, all the three combinations are generated again. We observed this and resolved in our study on such observed heterogeneity during ribosomal DNA ITS region amplification in Asiatic Vigna species (see Saini et al., Genet. Res., Camb. (2008), 90, pp. 299–316.). We also proved the differences (indel lengths, 2 bp and above) among clones by doing heteroduplex analysis by mixing different clones, in that study.

If you are interested in getting the amplicons, instead of band purification go for cloning and sequence.

all the best

Hydroethanol (30:70) at 40⁰C, assisted by shaking and or microwave assisted for total extractions.

Dear, Dr. Jessika Ferreira Nunes

It is an interesting question.

There will be a strong correlation between the two mentioned components (MWRM and ESG) because:

1- Environmental, Social and Governance (ESG) components can create a sustainable global solidarity ring.

2- Productivity management of raw material waste in industries reduces the need for natural resources and helps the sustainability of the environment.

3- The development of environmental sustainability is mostly operationalized in the shadow of three important aspects (ESG).

For more information on environmental sustainability, refer to the following link:

Greetings! You could try the following: Simple boiling. Employing chelex-100. Ultra pure water. Enzymic digestion: lysozyme. Heat and sodium hydroxide. Good luck!

Strategies to mitigate these challenges... Hope you can use my thoughts..🙂

There are several strategies in my mind....

Sample Dilution: The concentration of inhibitors can be reduced by diluting the sample as attempted. However, this approach may also dilute the target DNA to below detectable levels. The efficacy of this method is contingent on finding an optimal balance between dilution and target DNA concentration.

Optimized Lysis Protocols: To ensure efficient lysis of phytoplankton cells while minimizing the release of inhibitory substances, it is essential to incorporate both physical (freeze-thaw cycles) and chemical lysis methods. To ensure efficient lysis of phytoplankton cells while minimizing the release of inhibitory substances, it is essential to incorporate both physical (freeze-thaw cycles) and chemical lysis methods. The lysis conditions, such as buffer composition and incubation times, should be adjusted accordingly.

Additionally, the use of inhibitor-resistant enzymes, such as certain commercially available DNA polymerases, can improve DNA yield and purity. Using such enzymes can improve amplification success rates in challenging samples significantly (Schrader et al., 2012).

DNA purification is crucial for enhancing DNA quality. DNA purification kits or methods designed to remove polysaccharides, proteins, and other potential inhibitors are recommended. Magnetic bead-based cleanup protocols, as mentioned, are effective but may require optimization for phytoplankton-rich samples.

Alternative extraction methods should also be considered. Considering alternative DNA extraction methods that include steps specifically designed to remove inhibitors can be beneficial. For example, the use of CTAB (cetyltrimethylammonium bromide) in the extraction buffer can help to remove polysaccharides (Doyle & Doyle, 1987).

Implementing qPCR with internal controls can aid in quantifying the extent of PCR inhibition and verifying the amplification efficiency. This method allows for the detection and correction of samples that exhibit significant inhibition prior to library preparation.

Regards,

Phil

Since you have them already frozen in liquid nitrogen you can probably skip the lyophilization step. Grind them in a ball mill to get a nice powder. As Kai points out, you'll probably need water in your extraction and definitely need it for your analysis if you use HPLC.

For my DNA extraction, the last stage of elution is usually done with 70% ethanol, 80% ethanol (washing step), centrifugation, air drying the column and then eluting with hot water into another tube by centrifugation.

we are working on colored cotton fabric dna extraction kit

Hi,

If you have a DNAse treatment step during the extraction process, verification of the purity of your RNA might be optional.

However, for best practice, it is always good to run an agarose gel that shows:

-Purity of RNA (no gDNA on the top of the gel)

-Quality of the RNA (2 discrete bands corresponding to rRNA), no smear of degradation.

The latest is especially important ave inconsistent qPCR results with low-quality RNA.

When I run RT-qPCR, I prefer to always use the same quantity (usually 1 ug of total RNA) rather than simply adding equal volumes of random quantities.

Good luck!

Hello, Tomaz!

We have used the PurePrep 96 for about a year now, if I can be of any assistance to you.

Christopher

Usually, the first extraction can remove most impurities, while the second extraction further improves the purity of the target substance. By repeatedly extracting, the target substance can be effectively separated and impurities removed, resulting in a pure extract.

Bradford assay is typically given in terms of absorbance units (AU) at a specific wavelength, commonly 595 nm.

To convert the Bradford assay result into a percentage of protein, you need to establish a standard curve using known concentrations of protein. In this, Plot a graph of absorbance (y-axis) versus protein concentration (x-axis) for the standard solutions (at least 5 known samples). Once you have the protein concentration of your know sample, you can calculate the percentage of protein.

Divide it by the total protein content (in mg or µg) of the sample.

Percentage protein=Protein concentration of sample / Total protein in sample×100

This might be a stupid question but is your new cell line from the same species?

Thank you Phil Käding ! I performed yesterday the reaction in an extremely dried system and the quantity of the brown impurities was less than the first time.

0.001 ppb, and 0.0001 ppb..these are not reachable concentrations for MS systems even if you apply enrichment steps using large-volume SPE I guess...

Since the question is very not clear to me I should indicate that You need to develop a method by proving the working limits of the extraction are in alignment with the range of detection concentrations. Meaning, that extraction efficiency between 10 to 50 ppm is not something that validates 0.1 ppb, 0.001 ppb, and 0.0001 ppb levels.

Hi, follow this guide:

Materials Required

Frozen milk at -18°C.

Refrigerator set at 4°C for controlled thawing.

Cold water bath for accelerated thawing.

Sterile containers for collecting thawed milk.

Laminar flow hood or clean area to prevent contamination.

Methodology

1. Preparation Phase

Assessment of Initial Quality: Before proceeding with the thawing process, evaluate the initial quality of the frozen milk, noting any signs of freezer burn or contamination.

Selection of Containers: Choose appropriate sterile containers to collect the thawed milk. The containers should be pre-cooled to prevent sudden temperature changes upon transfer of milk.

2. Thawing Process

The thawing process is crucial and can be approached in two ways, depending on the urgency and the volume of milk to be thawed.Refrigerator Thawing (Slow Thawing Method):Transfer the frozen milk from the freezer (-18°C) to the refrigerator (4°C). This method is preferred for its gentle effect on milk components, reducing the risk of protein denaturation.Allow the milk to thaw gradually, which may take 24 to 48 hours, depending on the volume. Regularly check the progress and gently agitate the container to promote uniform thawing.Cold Water Bath Thawing (Accelerated Thawing Method):Place the frozen milk in a leak-proof bag if not already packaged accordingly.Submerge the bag in a cold water bath (not exceeding 10°C) to accelerate thawing while still preventing rapid temperature increases that could lead to protein damage.Change the water every 30 minutes to maintain a consistent thawing temperature.Once thawed, immediately transfer the milk to the refrigerator (4°C) to prevent microbial growth.

3. Post-Thaw Handling

Aseptic Handling: Conduct all transfers and handling of thawed milk in a laminar flow hood or clean area to prevent contamination.Quality Assessment: After thawing, assess the milk for any changes in physical appearance, smell, or consistency that may indicate degradation or contamination.Preparation for Lactoferrin Extraction: Proceed with lactoferrin extraction as soon as possible after thawing to minimize the time for potential microbial growth and further degradation of milk components.

That is Santolina africana. For sure. Abdelhak Maghchiche

Yes, i know. I already did in other biological matrices, like blood and bone marrow. But, we wanted to bring up some challenge and try to do PCR in serum samples. We have some ideas, but i just wanted to see if there was a different perspective.

Faizan Faraz Thank u

I found using Qiagen column could cause DNA/RNA breakage. Using original chemical method would be better for longer DNA rather than using column.

(late reply but hope this may help anyone come across the same issue)

I have similar experience with Qiagen plasmid prep kit, but I realise my precipitate was due to the LyseBlue added to the P1 buffer. An unopened bottle of P1 with no added LyseBlue should be clear.

In my experience, "Jelly" is almost aways caused by long, genomic DNA, with the warning that I have not worked with chromatic-associated RNA myself yet, I could imagine you could solve your problem by adding RNAse-free DNAse, there are commercial RNASe-free DNAse preparations available. I know it feels a bit scary to digest (fairly stable) DNA to isolate (fairly unstable)RNA. But the HBV group in my thesis lab did this routinely to get rid of plasmid DNA from transfection of HBV genomes. Worked for them to do HBV northern blots. To avoid the jelly, harsher pipetting (Yellow tips) might help too as it shears long DNA. but as a "clean" solution, the DNAse is probably better.

Good luck and let me know how that turns out as down the line I may have similar problems..

Best, Thomas :-)

Can Kiessling yeah, I am making a fresh Beta-mercaptoethanol.

I recently did a couple of minion runs. I used the Circulomics kit for size exclusion of fragments less than 25kb. I had a good N50 ranging from 12kb to 22kb with 16-20GB of data in one run. The only thing I suggest is not to freeze and thaw your DNA. If possible, perform the DNA extraction and library preparation on the same day; if not, don't freeze the DNA (you may use 4 degree storage but don't go higher). I think it's better to have more DNA than recommended during library preparation. For higher data, make sure your flow cells are within warranty, with a standard number of pores available for sequencing after the loading library.

Maybe Lukas Kleine could recommend you some other method?

The choice of the best organic solvent for solubility and extraction of nitrophenols depends on several factors, including the specific nitrophenol compound, the desired extraction efficiency, and the intended application. However, some commonly used organic solvents for the solubility and extraction of nitrophenols include:

The choice of solvent should be based on the specific requirements of your extraction process, including the polarity of the nitrophenol compound, the desired selectivity, and the safety considerations associated with the solvent.

Dear Nishitha Landgey ! It is necessary to destroy bacterial cells with a tissue shredder or ultrasound, and then carry out centrifugation. You will get a solution of carbohydrates in water.

To convert from ng/mL to mg/kg for a single extraction of heavy metal samples, you would need to take into account the density of the solution in which the heavy metals are present. Assuming a density of 1 g/mL for the solution:

It's important to be accurate with the measurements and units to ensure the conversion is done correctly.

It's commendable that you're trying to optimize your protocols for the separation of nucleus and cytoplasm in different cell types. Since you've had success with HeLa cells but are encountering sensitivity issues with SH-SY5Y cells, here are some suggestions to consider:

By systematically optimizing your protocol and considering the specific characteristics of SH-SY5Y cells, you can improve the efficiency and sensitivity of nucleus-cytoplasm separation for downstream applications.

The adaptation of wavelet transforms to the neural networks design used for classification enhances the feature extraction procedure by producing features invariant to different deformations of the images. In this respect, Stephan Mallat and Joan Bruna have conceived the scattering convolution network concept, see for example: Joan Bruna and Stephane Mallat, Invariant Scattering Convolution Networks, IEEE TRANSACTIONS ON PATTERN ANALYSIS AND MACHINE INTELLIGENCE, VOL. 35, NO. 8, AUGUST 2013. A wavelet scattering network computes a translation invariant image representation which is stable to deformations and preserves high-frequency information for classification. It cascades wavelet transform convolutions with nonlinear modulus and averaging operators. Recently, a Matlab toolbox, entitled Image scattering toolbox, was conceived.

Hi Mj,

it sounds like you want to use some sort of metabolomics software, which will produce an aligned peak list with integrals for each peak in each sample. these software normally align each peak as part of the analysis. I've never used metabolomics software to analyse masses which I've already extracted, only ever on the raw files from my MS (or converted to mzml), so im not sure if you need to change other parts of your workflow. I am also unaware of whether metabolomics software can handle MALDI output. But you have many options available, like MSDial, skyline, XCMS, the list goes on.

Hey Sajid Alvi I´m currently facing a similar problem , how did you solve it?

Thanks

Hey there Muhammad Tanveer! So, when it comes to extracting both organic carbon (OC) and elemental carbon (EC) from soot for measuring the band gap using the Tauc plot method, you'll want to choose a solvent that effectively dissolves these components. One commonly used solvent for this purpose is a mixture of toluene and acetone.

Toluene is great for dissolving organic components, including OC, while acetone is effective in extracting elemental carbon. The combination of these two solvents provides a comprehensive approach to extracting a wide range of carbonaceous materials from soot.

Here's a suggested procedure:

1. **Prepare the Solvent Mixture:**

- Mix toluene and acetone in a suitable ratio. A common ratio is 1:1, but you Muhammad Tanveer can adjust it based on your specific requirements.

2. **Soot Extraction:**

- Immerse the soot sample in the solvent mixture and let it soak for a sufficient period, ensuring thorough extraction.

3. **Separation:**

- After extraction, you Muhammad Tanveer can use filtration or centrifugation to separate the dissolved components from the insoluble residue.

4. **Analysis:**

- The extracted OC and EC can then be analyzed further for band gap measurements using the Tauc plot method.

Remember, the choice of solvent and its ratio may vary based on the composition of the soot and the specific properties you're aiming to study. Always conduct preliminary tests to optimize the solvent system for your particular soot sample.

While I am very excited to learn more from your research efforts, some of my work on OC and EC might be interesting for you Muhammad Tanveer to read:

Feel free to hit me up if you Muhammad Tanveer need more details or run into any hurdles along the way. Let's rock this experiment!

The best is freezing :)

I am aware that the question has been posed in 2018 I want to give my opinion anyway.

If the goal is to isolate mitochondrial DNA with minimal contamination of nuclear DNA then you should start with platelets (thrombocytes). These blood cells contain mitochondria but have no nucleus - at least in mammals.

So the first step should be to centrifuge the whole blood with an anticoagulant (EDTA). The red blood cells (without mitos) end up at the bottom, the buffy coat contains white blood cells (with lots of mitos but also with nuclei) and on top you have the serum with the platelets (mitos but no nuclei).

you can extract it in DEE before dissolving it in water.

Contact the manufacturer for the best advice regarding the product.

Multiple freeze-thaw cycles can cause the RNA in your samples to degrade. If the RNA is already degraded, then no extraction method will work. For your TriZol-extracted samples, are you able to see 28S and 18S bands? The 28S:18S ratio should be approximately 2:1.

Use Insoluble Polyvinypyrolidone in the extraction buffer

Freeze them with/in liquid nitrogen and/or putting in a -80 deg Celsius freezer for preventing lipid oxidation.

Often, in traditional methods heat is additionally applied. This might lead to chemical transformations of the compounds of interest.

Pigments are phytochemicals...

You can try and pre-extract (wash) your sample with solvents that will extract the pigments but not your constitute.

Either way 100mg/ml is a very high concentration to begin with. What are you extracting with?

At present and in the recent decade, one of the best available methods for extracting natural bioactive compounds and secondary metabolites from genuine/original germplasms is the enzyme-assisted-extraction method. Of course, This method has a very high costs.

https://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-biology/enzyme-assisted-extraction#:~:text=EAE%20works%20on%20the%20principle,to%20release%20the%20intracellular%20components.

Regards

Looks you want RNA from outer layer of kidney. Is it not possible for you to do microdissection under microscope and dissect the ourer layer. I will also recommend to do microdissection when whole kidney is under RNALATER solution, that will protect from RNAases

Dear friend Sarah L Fox

Hey there Sarah L Fox! I am here, ready to tackle your Western blot challenges. Now, histone modifications on H3 can indeed make it a tricky loading control. If H3 and H4 are not cutting it and Ponceau isn't playing nice, let's think outside the PVDF membrane!

Considering the inconsistencies with H4, lamin B might be worth a shot. Lamin B is a nuclear envelope protein and could serve as an alternative loading control, especially if you're working with nuclear extracts.

Here's a quick guide:

1. **Optimize Extraction:**

- Ensure your histone extraction is top-notch using the abcam kit. Proper extraction can save you Sarah L Fox from loading control woes.

2. **Membrane Type:**

- Since Ponceau isn't doing the trick, consider trying a nitrocellulose membrane. Sometimes, different membranes have different affinities.

3. **Lamin B as a Control:**

- Lamin B has been used as a loading control in some cases, especially in studies involving nuclear proteins. It's worth trying, but always validate with literature or pilot experiments.

4. **PVDF Stripping Woes:**

- If you're having trouble stripping the PVDF membrane, consider trying a fresh one for a cleaner start.

5. **Alternate Controls:**

- If lamin B doesn’t pan out, consider beta-actin or GAPDH. While they're not without their own challenges, they are commonly used and might suit your needs.

6. **Trial and Error:**

- Sometimes, it's all about experimenting. Tweak concentrations, try different gels, and play with transfer conditions. The perfect loading control might just be waiting for you Sarah L Fox to discover it.

7. **Consult the Literature:**

- Before you Sarah L Fox dive too deep, check recent literature. Others in your field might have encountered similar challenges and found solutions.

Remember, science is an adventure, and the best protocols often come from a bit of exploration. Let me know how it goes, and we'll navigate this Western blot wilderness together Sarah L Fox!

Dear Kashif Ali Please do recommend my answer if helpful

Solvent extraction can be a useful technique for purifying nitrogen-doped carbon quantum dots. Here's a general procedure you can follow:

**Materials Needed:**

1. Nitrogen-doped carbon quantum dots solution

2. Solvent (e.g., ethanol, acetone, or a suitable organic solvent)

3. Centrifuge

4. Filtration setup (e.g., syringe filter or filter paper)

5. Vacuum pump (if using a filtration setup)

6. Glassware and lab equipment

**Procedure:**

1. **Prepare Nitrogen-Doped Carbon Quantum Dots Solution:**

- Start with a solution of nitrogen-doped carbon quantum dots that may contain impurities or unreacted precursors.

2. **Selection of Solvent:**

- Choose a solvent that is suitable for extracting impurities or unreacted precursors while maintaining the stability of nitrogen-doped carbon quantum dots. Common solvents include ethanol or acetone.

3. **Extraction:**

- Mix the nitrogen-doped carbon quantum dots solution with the selected solvent. The impurities may preferentially dissolve in the solvent while the quantum dots remain in the solution.

4. **Centrifugation:**

- Centrifuge the mixture at a high speed to separate the impurities in the solvent from the quantum dots. The centrifugation time and speed depend on the specific characteristics of your solution.

5. **Decanting or Removing Supernatant:**

- Carefully decant or remove the supernatant (top layer) containing the dissolved impurities. Be cautious not to disturb the quantum dots pellet at the bottom.

6. **Washing (Optional):**

- Wash the quantum dots pellet with fresh solvent to remove any remaining impurities. Repeat the centrifugation and decanting process.

7. **Filtration:**

- If needed, filter the quantum dots solution using a syringe filter or filter paper to further remove smaller impurities or aggregates. Apply vacuum filtration if using filter paper.

8. **Characterization:**

- Characterize the purified nitrogen-doped carbon quantum dots using analytical techniques such as UV-Vis spectroscopy, fluorescence spectroscopy, or other characterization methods to confirm their purity and properties.

9. **Storage:**

- Store the purified nitrogen-doped carbon quantum dots in a suitable container, protecting them from light and contaminants.

**Notes:**

- Adjust the solvent choice and extraction conditions based on the properties of your nitrogen-doped carbon quantum dots.

- Use high-purity solvents to avoid introducing additional impurities.

- The purification process may need optimization based on the specific characteristics of your quantum dots and the impurities present.

Always follow safety guidelines and use appropriate protective equipment when working with chemicals and laboratory equipment. This procedure provides a general guideline, and you may need to adapt it based on your specific experimental conditions and requirements.

Mechanical separator can be used as an effective method for separation of microplastics from solid material.

What's this underselling yourself about? Taking a lesson and learning from it. That's the best you can ever do. I think that's exactly what you get full credit for.

Yes, there are other alternatives such as magnet bead-based methods or vacuum suckling devices when opting for silica columns-based methods.

Pandiyan Mu

Some steps you can follow for feature extraction:

1. Preprocessing: Clean and preprocess your data to remove any noise or artifacts that may interfere with the feature extraction process. This may involve filtering, normalization, or resampling the data.

2. Windowing: Split your data into smaller windows or segments. This is done to capture local characteristics and ensure that the statistical features are computed on meaningful subsets of the data. The window size will depend on the characteristics of your data and the expected fault frequencies.

3. Statistical Features: Compute various statistical features from each windowed segment. Some commonly used statistical features for vibration and acoustic data include:

- Mean: Average value of the data within each window.

- Standard Deviation: Measure of the dispersion of the data within each window.

- RMS (Root Mean Square): Square root of the average of the squared values within each window, providing information about the signal's energy.

- Kurtosis: Measure of the peakedness or flatness of the data distribution within each window.

- Skewness: Measure of the asymmetry of the data distribution within each window.

- Crest Factor: Ratio of the peak value to the RMS value within each window, indicating the signal's peakiness.

Additionally, you can consider other statistical features such as minimum, maximum, range, variance, and percentile values.

4. Feature Selection: After computing the statistical features, you may want to perform feature selection to identify the most relevant and informative features for your ML model. This can help reduce dimensionality and improve the model's performance. Techniques like correlation analysis, feature importance ranking, or domain knowledge can be used for feature selection.

Once you have extracted the relevant features, you can use them as input to train your ML algorithm.

Good luck: partial credit AI

A lipid extraction procedure quantitatively extracts cellular lipids in an undegraded state and uncontaminated with nonlipid components such as free sugars and amino acids. The two most conventional methods of lipid extraction are, namely, Folch method and Bligh and Dyer method.Enveloped viruses acquire lipid membranes as their outer coat through interactions with cellular membranes during morphogenesis within, and egress from, infected cells. In contrast, non-enveloped viruses typically exit cells by cell lysis, and lipid membranes are not part of the released virions. Viral envelopes consist of a lipid bilayer that closely surrounds a shell of virus-encoded membrane-associated proteins. The exterior of the bilayer is studded with virus-coded, glycosylated (trans-) membrane proteins. The main component of the viral envelope is the host-derived lipid bilayer. The precise composition of this lipid membrane varies, as different viruses acquire their envelopes from different cellular membranes. The virus envelope is known as a capsid. Capsid protects the genetic material of the virus during the entire life cycle of the virus.these issues, the multi-omics sample preparation technique MPLEx (metabolite, protein, and lipid extraction) is developed to partition a single sample into three distinct parts (metabolites, proteins, and lipids) for multi-omics analysis, while simultaneously inactivating MERS-CoV by solubilizing and disrupting the viral envelope and denaturing viral proteins.

Hello Arsiema Berhe

If your DNA extraction mix is 1000ul, then you will add 10ul of (100X) Triton X-100 to give you (1X) Triton X-100 in final DNA extraction mix.

Similarly, if your DNA extraction mix is 500ul, then you will add 5ul of (100X) Triton X-100 to give you (1X) Triton X-100 in final DNA extraction mix.

So, you need not dilute (100X)Triton X-100.

Best.

Dear friend Saeede Taherpanah

Now, let's dive into the world of mycosporin-like amino acids (MAAs) with my fervor!

For a quick method of extracting MAAs from microalgae with a primary measurement using spectrophotometry, you Saeede Taherpanah can consider a simple and rapid acidified extraction method. Here's a brief guide:

### Quick MAA Extraction Method:

**Materials Needed:**

- Microalgae samples

- Methanol

- Acid (e.g., hydrochloric acid)

- Water

- Spectrophotometer

**Procedure:**

1. **Harvest Microalgae:**

- Collect microalgae samples during the exponential growth phase.

refer:

2. **Sample Preparation:**

- Wash microalgae with distilled water to remove salts.

- Freeze the samples and lyophilize to remove excess water.

3. **Acidified Methanol Extraction:**

- Add a known quantity of microalgae powder to a vial.

- Add methanol (typically 80-100%) acidified with a small amount of hydrochloric acid (0.1-1%) to the vial.

- Shake vigorously for a few minutes.

4. **Centrifugation:**

- Centrifuge the mixture to separate the cellular debris from the extract.

5. **Spectrophotometric Measurement:**

- Take a small aliquot of the supernatant.

- Measure the absorbance at a wavelength specific to the type of MAA present (typically around 310-360 nm).

6. **Calculation:**

- Use the Beer-Lambert Law to calculate the MAA concentration:

A=ε⋅c⋅l

where

- A is the absorbance,

- ε is the molar absorptivity of the specific MAA,

- c is the concentration, and

- l is the path length.

**Notes:**

- Calibration curves with known MAA standards can help in quantification.

- The choice of acid and its concentration can influence the extraction efficiency.

This method provides a rapid way to screen strains for the presence of MAAs using spectrophotometry. Keep in mind that it might not give the same precision as HPLC, but it's a trade-off for speed.

Now, go forth and unravel the mysteries of microalgae with my spirit!

Various methods, including ultrasound, supercritical fluid (SFE), microwave, and subcritical CO2-assisted techniques, have been investigated for extracting bioactive compounds from diverse plant matrices. These approaches provide alternative solutions to tackle challenges posed by interfering particles during the extraction process.

Moreover, it is crucial to take into account the influence of changes in ion concentration on the extraction process. For instance, modifying the pH of the medium using diluted HCl or NaOH can alter the ion concentration, potentially leading to adverse effects on the extraction of bioactive compounds.

Increasing RNA yield from mice peritoneal macrophages during the extraction process involves optimizing various steps to enhance the efficiency of RNA isolation. Here's a detailed guide on how to achieve higher RNA yields:

1. Cell Culture Optimization:

2. Harvesting Cells:

3. Cell Lysis:

4. Homogenization:

5. RNA Stabilization:

6. RNA Extraction Kit Selection:

7. DNase Treatment:

8. Column Washes:

9. Elution Volume:

10. Quantification and Quality Assessment:

11. Repeat Extractions if Necessary:

12. Temperature Control:

13. Standardize Protocols:

14. Troubleshooting:

15. Collaborate and Seek Advice:

By systematically optimizing each step of the RNA extraction process, you can enhance the yield and quality of RNA extracted from mice peritoneal macrophages. Always validate the extracted RNA through appropriate quantification and quality assessment methods before proceeding to downstream applications.

hello. in the following site you will find the desired information:

What do you propose to extract?

The miniprep from high manipulated cell type like Top 10 and high copy number pls like pCMV from 3-5 ml preculture should give around 400-700 ng/ul pls.

Hi Richard,

Thank you for replying to the question. Can you please share any relevant publications or documents where I can review this method?

Regarding the magnetic surface group, it's essential to select magnetic beads that are functionalized with ligands or molecules suitable for your application. The specific group or ligand on the magnetic surface should interact selectively with your target molecules. Common functional groups include streptavidin, protein A/G, and various antibodies. The choice of the group depends on the molecules you are trying to capture or separate.

When troubleshooting your protocol, it's a good idea to systematically examine each step, considering factors like buffer pH and composition, temperature, and the specific properties of your magnetic beads. Identifying the root cause of any issues can help you refine your protocol for better results

Check out these articles.

Thanks

Researchers should cite paper that 1. It has been published in reliable scientific journals. 2. In connection with the research that has been done. 3. Valuable articles in terms of content, for example review articles. 4. It should have a high ranking and citation, which shows the value and practicality of the article.

Well...I must say that this is not due to phenol/chloroform. This is a very common extraction method. What I assume is that after RNA precipitation, one should be careful to avoid contamination of DNA/protein that is frequently pipetted during RNA collection. I recommend for re-experiment in a sterile environment with careful harvesting of RNA. Best wishes.