Im working with the Gecko v2 (1-vector system) library and followed the Joung et al. (2017) Nature Protocols paper very closely.
Now i'm in the screening section and organizing next PCR for NGS step.
Then, i have questions.
1. If i am followed the protocol, i'll need 10 different forward primers. Why cannot i use single forward primer??
I'm hesitating to use all of them because it's too expensive to prepare 10 primers ( w/ 100 bases ).
2. Instead of 1 step PCR, can i try with 2 step PCR (done in 2014, science) because it's cheaper than using w/ 10 primers.