I did extraction for a DNA from blood on a filter paper using phenol chloroform method and got a purity for many samples less than 1.8 and low concentration. I used qiagen kit for the concerned samples and got more or less the same result. Any suggestions of how to solve this problem?
I would suggest you the DNA pellet which you got after extraction resuspend it phenol chloroform and isoamyl alcohol solution and again try to get the dna pellet then check for the purity repeat the same steps for 2-3 times and you will get pure DNA.
Otherwise use CTAB method for DNA extraction.
I hope this will help you
I would agree with Surjit, a second phenol chloroform extraction will help remove any protein contaminants. I would also suggest that if you precipitate your sample following phenol extraction it's good to perform 2 washes with 80% ethanol to help remove any phenol that can affect the 260/230 ratio and potential downstream experiments - PCR etc.
Good luck
You may treat suspended DNA with Proteinase k followed by phenol chloroform
I will try proteinase k then phenol chloroform isoamply and finally washing.Hope it works.
Dear Rihab
Plz read page 436-441 of this book for better understanding of contamination Issues.
This book will help you a lot in the life to come.
Best Regards
Hi,
Thank you tremendously for this book.I opened it and it is a very good book.Thank you again for your help.
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