genomic DNA yield

[Meenu Kesarwani]

Hi Julia and the community

I am using Zymo midi plus kit as suggested in the protocol and did 2 columns/ 50M cells. I got very low yield 70ng/ul total of 600ul ( I did two elution, first one with 200 ul and the second with 100). I am wondering what is typical yield with this kit?

Also for the PCR amplification how much can i / reaction of 50 ul use since it is so dilute. and is it enough DNA and will it represent the library well?

Please let me know any info will be helpful going forward.

Thank you

Meenu

[Tanja Müller]

Hello Meenu,

I am facing a similar issue and was wondering whether you have meanwhile proceeded with NGS or not?

I used the same gDNA extraction kit from Zymo. My gDNA yield of 4 samples (each 50Mio cells) was for all the samples around +/- 50µg total gDNA. Binding capacity of the Zymo Midi column should be 125µg and I used 2x columns per sample.

I repeated the gRNA extraction with 150Mio cells and obtained 170µg. I do not know how others get >200µg for 50Mio cells... Maybe my cell counter is not working correctly :D

Would be interested in hearing whether you managed to get good NGS results eventhough gDNA seems to be not enough.

Best wishes,

Tanja