For many years I've been using a simple solution of guanidine hydrochloride and water as my lysis buffer when doing DNA extraction (both for arthropods and fungi). It's always worked fine for me and it's super fast (10 minutes at 65°C) and cheap (about 15 cents per extraction). However, these days pretty much everyone uses a Proteinase K-based lysis buffer (often with a detergent as well). However, using Proteinase K is slow (typically a few hours or overnight) and more expensive.
What are the advantages of using Proteinase K? Is it really worth the added time and expense? (My end use is Sanger sequencing the DNA.)
Hi Valerie,
the main advantage of using Proteinase K over GuHCl is that GuHCl can potentially interfere with downstream applications if not completely removed during the DNA extraction process. Residual guanidine hydrochloride can inhibit enzymatic reactions such as PCR (polymerase chain reaction) and may affect the quality and integrity of the extracted DNA. Therefore, thorough washing and purification steps are necessary to remove any traces of guanidine hydrochloride. Whereas removal or inactivation of Proteinase K is much easier.
Best,
Murat
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