I need to recover randomly mutagenized plasmids and sequence for the mutation from each hit of a screen yielding ~100 hits. The two options I am considering are (1) a yeast plasmid isolation procedure involving glass bead lysis and boiling followed by transforming recovered DNA into competent E coli or (2) colony PCR with universal primers across the mutagenized region followed by exonuclease and SAP treatment of reactions (destroys primers and dNTPs) followed by sequencing of amplicon. It makes no difference to me whether the saved glycerol stock of clones is in E coli or yeast cells. I'm looking for speed of sequencing and cost-effectiveness.
- Badges
- Science topic
Similar questions and discussions