Hello!
I have submitted genomic DNA for WGS to our core sequencing facility. The nanodrop reading says my DNA is ~50ng/microliter, but the pico green (which I am told is specific for dsDNA) is much lower. The core facility tech suggests that my DNA may be ssDNA, but I don't know why it would be ssDNA. I prepared the DNA using phenol chloroform extraction followed by EtOH precipitation. The DNA was resuspended in molecular grade water at 4 degrees for 24 hrs. The DNA looks good on a gel (it runs at >12kb fragments) and DNA prepared with the same protocol is cut with restriction enzymes (which cut only dsDNA).
I don't know what the problem with my DNA is and if I should tell the technician to proceed to library prep with my samples or not.
Please help me anonymous smart people!
Thanks
SaraH
I have submitted genomic DNA for WGS to our core sequencing facility. The nanodrop reading says my DNA is ~50ng/microliter, but the pico green (which I am told is specific for dsDNA) is much lower. The core facility tech suggests that my DNA may be ssDNA, but I don't know why it would be ssDNA. I prepared the DNA using phenol chloroform extraction followed by EtOH precipitation. The DNA was resuspended in molecular grade water at 4 degrees for 24 hrs. The DNA looks good on a gel (it runs at >12kb fragments) and DNA prepared with the same protocol is cut with restriction enzymes (which cut only dsDNA).
I don't know what the problem with my DNA is and if I should tell the technician to proceed to library prep with my samples or not.
Please help me anonymous smart people!
Thanks
SaraH
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