Can you help what must I do to purify total RNA from klebsiella pneumonaie? I have GeneJet RNA purification kit and it does not include lysozyme. It contains proteinase k. Manufacturer protocol said RNA purification from bacteria, you must use lysozyme. Can I use proteinase k instead of lysozyme or lysozyme requires for purification?
Hi there,
If you intend to extract RNA from bacteria, you'd better use lysozyme which digests peptidoglycan in order to lyse cells. Proteinase K replacing lysozyme will be useless.
hii
lysozyme mostly will be using during plasmid, rna extractions from bacteria for the cell lysis. Proteinase K plays the role for protein degardation.
Both the chemicals are having different roles.
Hello, on some processes based on protein degrading you can use lysozyme or proteinase K or both together, but as I understand from your situation it is very better to use lysozyme rather than P-K.
Dear Fatih and Raksha
First of all thanks for your interest. i will try to find lysozyme.
Best regards
Hi İbrahim I used to Proteinase K for total nucleic acid extraction from both bacteria and viruses, and proteinase K was supplied by the manufacturer's kit. My opinion is that you should try Proteinase K and you should make some modifications on the durations of procedure steps for example you should incubate the bacteria with proteinase K for 15 min instead of 10 min at 50 C. Good luck
Dear Fatih, I'm coming back on this post since I have a question concerning your comment. You say that you can use both lysozyme and proteinase K at the same time, but isn't the proteinase k going to diggest the lysozyme ?
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