I am looking to do targeted sequencing on approximately ~40Mb of mouse genome (~2% of the genome) from low input DNA (~100pg). Equivalent to sequencing the equivalent of 2 exomes worth of DNA from 20 cells.
My current approach has been to design large primer pools to PCR amplify ~40Mb from ~100pg DNA input thus capturing and amplifying the required 40Mb in one step. I have had varying levels of success with this approach but I am now looking for alternative methods.
Has anyone been doing a similar sort of protocol? Or maybe have any ideas on how I could reproducibly capture and amplify 2% of a mouse genome from low DNA input?
Thanks!
My current approach has been to design large primer pools to PCR amplify ~40Mb from ~100pg DNA input thus capturing and amplifying the required 40Mb in one step. I have had varying levels of success with this approach but I am now looking for alternative methods.
Has anyone been doing a similar sort of protocol? Or maybe have any ideas on how I could reproducibly capture and amplify 2% of a mouse genome from low DNA input?
Thanks!
Comment