I'm making nucleic acid extraction with Maxwell RSC equipment. But the purity 260/280 is not too high. I extract the nucleic acids from plasma samples. So I'd like to know if there's anyway to improve purity. I make quantification with Nanodrop 200.
Regards.
Dear Sir. Concerning your issue about the purification of the DNA/RNA Extracts. Extraction of DNA, RNA, and protein is the basic method used in molecular biology. These biomolecules can be isolated from any biological material for subsequent downstream processes, analytical, or preparative purposes. In the past, the process of extraction and purification of nucleic acids used to be complicated, time-consuming, labor-intensive, and limited in terms of overall throughput. Currently, there are many specialized methods that can be used to extract pure biomolecules, such as solution-based and column-based protocols. Manual method has certainly come a long way over time with various commercial offerings which included complete kits containing most of the components needed to isolate nucleic acid, but most of them require repeated centrifugation steps, followed by removal of supernatants depending on the type of specimen and additional mechanical treatment. Automated systems designed for medium-to-large laboratories have grown in demand over recent years. It is an alternative to labor-intensive manual methods. The technology should allow a high throughput of samples; the yield, purity, reproducibility, and scalability of the biomolecules as well as the speed, accuracy, and reliability of the assay should be maximal, while minimizing the risk of cross-contamination. Extraction of DNA, RNA, and protein is the basic method used in molecular biology. These biomolecules can be isolated from any biological material for subsequent downstream processes, analytical, or preparative purposes. In the past, the process of extraction and purification of nucleic acids used to be complicated, time-consuming, labor-intensive, and limited in terms of overall throughput. Currently, there are many specialized methods that can be used to extract pure biomolecules, such as solution-based and column-based protocols. Manual method has certainly come a long way over time with various commercial offerings which included complete kits containing most of the components needed to isolate nucleic acid, but most of them require repeated centrifugation steps, followed by removal of supernatants depending on the type of specimen and additional mechanical treatment. Automated systems designed for medium-to-large laboratories have grown in demand over recent years. It is an alternative to labor-intensive manual methods. The technology should allow a high throughput of samples; the yield, purity, reproducibility, and scalability of the biomolecules as well as the speed, accuracy, and reliability of the assay should be maximal, while minimizing the risk of cross-contamination. Extraction of DNA, RNA, and protein is the basic method used in molecular biology. These biomolecules can be isolated from any biological material for subsequent downstream processes, analytical, or preparative purposes. In the past, the process of extraction and purification of nucleic acids used to be complicated, time-consuming, labor-intensive, and limited in terms of overall throughput. Currently, there are many specialized methods that can be used to extract pure biomolecules, such as solution-based and column-based protocols. Manual method has certainly come a long way over time with various commercial offerings which included complete kits containing most of the components needed to isolate nucleic acid, but most of them require repeated centrifugation steps, followed by removal of supernatants depending on the type of specimen and additional mechanical treatment. Automated systems designed for medium-to-large laboratories have grown in demand over recent years. It is an alternative to labor-intensive manual methods. The technology should allow a high throughput of samples; the yield, purity, reproducibility, and scalability of the biomolecules as well as the speed, accuracy, and reliability of the assay should be maximal, while minimizing the risk of cross-contamination. I think the following below links may help you in your analysis:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2789530/pdf/JBB2009-574398.pdf https://info.gbiosciences.com/blog/bid/177600/dna-purification-vs-rna-purification-what-is-the-difference
http://www.tataa.com/core-facility/laboratory-areas/extraction-and-purification-of-rna-mirna-dna-and-protein/
Thanks
When I worked by qRT-PCR, I had similar problem for extraction of RNA. I used Total RNA isolation - Macherey Nagel for my purpose. there were two lysate buffers, I checked both of them and I saw one of them was better, and additionally in the stage of adding DNase to the column, instead of one time and 15 min, I used two times and 30 min.
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Briefly I think you must use the best isolation kit. and you must give more attention to contamination, and also on preservation of samples.
thanks
I must say the nano drop is not a very reliable way of measuring your RNA concentration and therefore integrity. Furthermore, in my experience you get the best quality RNA and the highest yield simply using Trizol. The samples I have collected using Trizol had much higher RNA integrity scores than RNA yielded by several commercial kits. If you would like to clean your samples you can try to reprecipitate your RNA and redo the cleanup. It worked very well for me, but in my case, the 260/230 ratio in a couple of samples was low. And I must say that depending on what you would like to do with the RNA, your contamination might not be such a problem.
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