RNA is insoluble in isopropanol so it will aggregate together, giving a pellet upon centrifugation. This step also removes alcohol-soluble salt.
Isopropanol is more preferred than ethanol in DNA extraction, as isopropanol facilitates precipitation more better, as it possess very less i.e., 0.6 to 0.7 volumes of alcohol.
Isopropanol precipitates the RNA. Up to that point it's generally in solution. Centrifuging the tube after this step should leave a very faint but generally visible white smudge/pellet of RNA. The ethanol steps that follow the isopropanol precipitation are simple washes.
RNAse destroys the RNA and hence RNAse contamination is a problem in RNA extraction as it breaks down RNA. RNAse enzyme is removed by using RNAse inhibitor or precautions like wearing of gloves, autoclaving tips , using RNAse free water/DEPC treated water is done while performing RTPCR
To concentrate or purify the DNA, which is insoluble in isopropanol. Once the solution containing your DNA is placed in isopropanol and centrifuged, the DNA will precipitate to a little pellet at the bottom of your tube. Everything else in your tube is soluble in isopropanol and will remain in liquid form. Pipet the liquid out and now you have just DNA.
Trizol is a common reagent used for RNA isolation from biological samples. It works by disrupting cells and denaturing proteins to release RNA. Trizol also aids in the separation of RNA from other cellular components, allowing for efficient and high-yield RNA extraction.
Isopropanol is used in DNA extraction to separate DNA from other cellular components. It helps to precipitate the DNA, causing it to clump together and separate from the rest of the solution. This allows for the isolation and purification of the DNA for further analysis.
Isopropanol is more preferred than ethanol in DNA extraction, as isopropanol facilitates precipitation more better, as it possess very less i.e., 0.6 to 0.7 volumes of alcohol.
I have not personally used the Qiagen Total RNA Extraction Kit for RNA extraction.
Chloroform is commonly used in RNA extraction to separate RNA from other cellular components. It helps in the denaturation of proteins and the dissolution of lipids during the extraction process. Chloroform aids in the formation of a distinct organic phase where RNA can be collected.
Isopropanol precipitates the RNA. Up to that point it's generally in solution. Centrifuging the tube after this step should leave a very faint but generally visible white smudge/pellet of RNA. The ethanol steps that follow the isopropanol precipitation are simple washes.
RNAse destroys the RNA and hence RNAse contamination is a problem in RNA extraction as it breaks down RNA. RNAse enzyme is removed by using RNAse inhibitor or precautions like wearing of gloves, autoclaving tips , using RNAse free water/DEPC treated water is done while performing RTPCR
"b -mercaptoethanol is used to help to destroy RNases that may be present and will degrade the RNA. b -mercaptoethanol is a reducing agent that will reduce the disulfide bonds of the RNases, thereby destroying the conformation and the functionality of the enzyme". It comes from http://www.norgenbiotek.com/index.php?id=faqs_rnakits
To concentrate or purify the DNA, which is insoluble in isopropanol. Once the solution containing your DNA is placed in isopropanol and centrifuged, the DNA will precipitate to a little pellet at the bottom of your tube. Everything else in your tube is soluble in isopropanol and will remain in liquid form. Pipet the liquid out and now you have just DNA.
TE stands for Tris and EDTA. The Tris buffers the water to prevent acid hydrolysis of the DNA/RNA. The EDTA chelates divalent cations that can assist in the degradation of RNA.
75% ethanol is commonly used in RNA extraction because it helps to wash the RNA pellet by removing salts and other contaminants, while also helping to maintain the integrity and stability of RNA molecules. The lower ethanol concentration reduces the risk of RNA degradation and allows for efficient RNA recovery during the extraction process.
Adjusting the pH to 7 during RNA extraction helps to create the optimal conditions for RNA stability. RNA is more stable at a neutral pH, which minimizes degradation and helps maintain the integrity of the RNA molecules during the extraction process. This ensures that high-quality RNA is obtained for downstream applications.
Seventy percent ethanol is commonly used in RNA extraction to wash and remove salts and contaminants from the RNA sample. It helps to purify the RNA by precipitating it out of the solution while leaving behind impurities. Additionally, the 70% ethanol concentration helps minimize RNA degradation during the extraction process.