Library titer for ZAP cDNA

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Library titer
for ZAP cDNA
  • Want the dilution that gives 10,000 plaques per 100 mm diameter plate
  • Keep all dilutions made from the library for later use


Day 1
  • Streak LE392 onto a NZY or LBM plate and grow o/n at 37°C
Day 2
  • Start an o/n from the fresh plate of LE392 in LB with 10mM MgSO4
  • Make the following dilution series:






  • Combine 10 l of dilution to be plated with 100 l of LE392 in 1.7 ml bullet
  • Warm (no shaking) at 37°C for 15 min
  • Add phage/LE392 to 3 ml lambda top agarose cooled to 50°C
  • Working quickly, vortex for 10 seconds and pour all 3 ml onto a pre-warmed (37°C) NZY or LBM plate; rock plate to spread agarose out over entire surface
  • Once plates are solidified, place at 37°C o/n
Day 3
  • Count plate(s) that show 30-300 plaques per plate
  • Back calculate to determine library concentration
  • Determine dilution to use for 10,000 plaques per plate

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