oligopaint probe making planning

emPCR ready libraries:

  • AbdA
  • AbdB
  • Ubx
  • en-locus 100kb
  • Black 100kb
  • Yellow 100kb
  • Green 100kb

Synthesis schemes:

Teritaries + Nicking

  • PCR using extended primers = Forward primer (21bp) + Secondary binding site (32 bp)
  • Nick off R primer
  • Denaturing PAGE Gel extract

Tertiaries + Exonuclease

  • Start with same PCR + extended primers
  • Second PCR using PPT-primer in place of extended primer
  • Exonuclease digestion
  • Denaturing PAGE Gel extract

Assymetric PCR

  • Start with same PCR + extended primers
  • Second PCR using just extended F primers, no reverse. 40 rounds?
  • Run on PAGE
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