Jasieniuk:Notebook/Apomixis in hybrid Rubus/2010/11/15

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PCR setup

  • RUB101115A = RUB262/CBA14 for S02, B, and L. Excel sheets set up by Stella 11/12 for RUB262 and CBA14 were NOT used since I would still like to multiplex RUB262 and CBA14 at this stage. DNA was pipetted onto a plate, and partial PCR mixes were made and frozen (see below).
  • RUB101115B = RUB126 for A, S01. This PCR was run today.
  • RUB101115C = RUB262/12 for parents, A, and S01. This PCR was run today.
  • For all three plates, samples were loaded into the same wells in which they will be on the ABI plate, according to the "do-overs 101112.xlsx" file. This differs from the PCR run 11/13, but I have a spreadsheet printed out showing how they will go together for the dilution. I will leave this on the bench in case you are in before me.

Two partial PCR mixes were made for RUB101115A and put into Stella's box.

  • The tube labeled "262 A" has 42.9 ul buffer, 17.1 ul dNTPs, and 7.5 ul RUB262 added to it. It needs 229.5 ul water and 3 ul Taq.
  • The tube labeled "262/14 A" has 57.2 ul buffer, 22.8 ul dNTPs, 4 ul CBA14, and 40 ul RUB262. It needs 272 ul water and 4 ul Taq.

Tasks for tomorrow:

  • Run PCR for RUB101115A. This involves adding water and Taq to the PCR tubes, pipetting PCR mix onto the plate according to the "do-overs 101112.xlsx" file, and putting it on the machine.
  • Dilution plates. All the PCR is done for the second plate (parents, A, and S01) so this dilution plate can be made if there is extra time after the PCR setup. Either way, Lindsay will make the first dilution plate (S02, B, L) in the afternoon after the PCR is done.
  • Formamide plates.


Stella: PCR

  • Run PCR of RUB262/CBA14 for S02, B, and L, using "do-overs 101112.xls" well information.
  • Changed entry of 11/12: deleted RUB262 and CBA14 excel file, keeping RUB126 excel file for information purposes.


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