Determining Phage Titer

  1. Microwave flask of Top Agar (LB + 0.8% agar) to melt. Allow to cool until handleable, then prepare 4 ml aliquots in small test tubes for as many samples as you will be handling. Place tubes in a heat block set at 55-60C.
  2. Prepare dilutions of the phage stock in LB. Typically a dilution series from 10<sup>0</sup> to 10<sup>10</sup> in 10<sup>2</sup> increments will give one plate with a countable number of plaques. This can be easily prepared by making 1.7 ml Eppendorf tubes with 990 l LB and transferring 10 l from the higher stock, mixing, and transferring to the next sample.
  3. Combine 100 l of exponentially growing host cells and 100 l of phage dilution. Incubate unshaken at 37C for 30 minutes.
  4. Add each sample to the top agar aliquot. Mix by pipetting up and down to avoid bubbles. Pour entire contents of test tube on top of an LB plate.

-- Main.ColinBrown - 14 Dec 2017

Edit | Attach | Watch | Print version | History: r6 | r4 < r3 < r2 < r1 | Backlinks | Raw View | More topic actions...

 Barrick Lab  >  ProtocolList  >  ProtocolsPhageAdsorptionRate  >  ProtocolPhageLysate  >  ProtocolsPhageTiters

Contributors to this topic Edit topic JeffreyBarrick, ColinBrown, CameronRoots
Topic revision: r1 - 2017-12-14 - 22:02:23 - Main.ColinBrown
 
This site is powered by the TWiki collaboration platform Powered by Perl This site is powered by the TWiki collaboration platformCopyright ©2022 Barrick Lab contributing authors. Ideas, requests, problems? Send feedback