Another perspective on the DNA contamination
Why no love for ssDNA?
Dr. Arakawa brings up an important point. ssDNA can also create havoc in the cell and shouldn’t we also be quantitating that?
This is a valid point. I guess we will need to blow the dust off those OliGreen kits and see how much DNA is ssDNA. We are using heat to open up the LNPs but the Cot curve for highly similar short DNA should be very fast. Cot curves measure the rate of reannealing DNA from ssDNA into dsDNA. It takes a long time for complex genomes but for small fragmented plasmids these timescales should be minutes.
An OliGreen measurement may push the numbers a little higher. I’ll see if we have a kit floating around. We will need to look over the regulations as they may speak to dsDNA contamination specifically. There certainly spend alot of time speaking about dsRNA just cant recall if they care about ssDNA vs dsDNA.
Next week I will presenting at NCI on these topics.
But they told us if i can remember right, that there is no DNA in the Vaccine means 0 zero not 1-5000 Tolerance range. That means any tolerance Or value greater than 0 is not acceptable!
Random frameshift question; How many frameshift mutations does the average human genome contain?