I'm looking for an alternative method for the extraction of genomic DNA from E. coli. Is there a simpler way other than using phenol:chloroform and extraction kit? I've found a method that was previously used for blood cells. Will it be the same if I were to use it for E. coli?
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC334765/pdf/nar00145-0424.pdf
I've found this one too, exactly for E. coli without using phenol:chloroform yet I was not able to get any original source out of it. How much can I trust this source? Has anyone found the source?
http://wikieducator.org/ANDC_DU/Biology_Protocols/Genomic_DNA
Thank you in advance for answering my question.
The link you posted is for an inorganic method, using TNES (Tris, NaCl, EDTA, SDS). I get very good results isolating genomic DNA using TNES for lysis, followed by protein precipitation, then isopropanol precipitation of DNA. "Salting out" refers to the precipitation of DNA using isopropanol in the presence of salt. If you're interested in doing it this way, I can give you my protocol.
Hi Wong,
Simplest method for isolating DNA is heat treatment without addition of any reagents.
1.Overnight bacterial colonies can be put in 1 mL of DI water in a test tube. Boil it for 10 minutes in a water bath and then centrifuge at 1000 rpm for 5 min. 5uL of supernatant can be used for PCR analysis.
2. you can take a few overnight bacterial colonies in 1mL of distilled water and heat in Microwave oven for 5-10 seconds followed by centrifugation at 1000 rpm for 2 min. 5uL of supernatant can be used or PCR analysis.
You can extract enough DNA by this method.
Good luck
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