Hi Alina,

I'm a little confused if you want to measure genome size or concentration? I use the Qubit all the time to measure genomic DNA concentration, but I don't know how you would use it for genome size. I've heard of people estimating relative genome sizes of plants using flow cytometry, but I don't have a specific protocol for you.

One tip when measuring gDNA concentration, is to make it reasonably dilute as it can become viscous when highly concentrated, making determination inaccurate.

Hope this helps,

Andrew

Andrew, thank you very much for your response! This is a good question. I've also from the literature gathered that we can use a DNA concentration as a proxy for genome size, and then, there is some formula to convert the result to the actual genome size. I'm hoping to get more insight on that..Thank you for your suggestion for measuring DNA concentration, I appreciate it.

Dear Alina,

I suggesting you few trouble shooting for.

1. Check your DNA concentration, quality over gel first

2. Quality of extracted DNA check with Nanodrop and plant specific Housekeeping Gene PCR

3. Measure concentration of total DNA on nanodrop or qubit and Serial Dilution of total genomic DNA up to femtogram.

4. Put standard curve PCR or detection limit of the target using PCR, based on detection limit, Amplicon Size, target Size, calculate number of copy using Avogadro number for actual quantification.

Best

Dear Subash, thank you for your suggestions - they are very helpful! I've done steps 1-2, I'll figure out the serial dilutions next. Thanks again!