SubmittingFam20COct

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October 14, 2014: DNA pick-up by [http://sequetech.com/ Sequetech] to sequence 2 samples, detailed below

  • Location: BioCurious
  • Participants: Rachel (set up sample pick-up), Emi (gave samples to Sequetech rep)
  • Notes: Rachel
  • Sequencing Strategy & Methods:
    • Details about samples submitted for sequencing in table, below.
    • Submitting 10uL aliquots of 2 samples: Sap.FAKS.hFam20C(Kex+).Sap:pD1214 and Sap.Faks.hFam20C(Kex-).Sap:pD1214 from Cloning bovine beta casein, Fam20C (Kex + & -), from 27Sep2014. Submitting clones 9A and 10A. Should have been 9B; 9A is "bad" per CR.
    • Sequetech rep. collected samples at ~2:00pm today.
    • All to be sequenced in three reactions: two forward primers (Fam20C For 640 & Fam20C For 1035) and one reverse (FAM20C Rev 1205). All primers designed 09Oct2014 by RVC project members.
Primer Sequences 
   
Fam20C For 640: 5' - CCTGCTTAGAGATCCA - 3' 
 Binds internally in Sap.Faks.hFam20C(Kex + & Kex-).Sap gBlock DNA and reads through in the 5'-3' direction.

Fam20C For 1035: 5' - ATTGATTATGACCTTCCA -3'
 Binds internally in Sap.Faks.hFam20C(Kex + & Kex-).Sap gBlock DNA downstream of Fam20C For 640 and reads     
 through in the 5'-3' direction.

FAM20C Rev 1205 : 5'-ATTCTACCAGCAACAG -3'
 Binds internally in Sap.Faks.hFam20C(Kex + & Kex-).Sap gBlock DNA and reads through in the 3'-5' direction.
 

Details for 6 sequencing reactions on plasmid DNA from 2 clones:

Sample details from Sequetech sequence pick-up, 14Oct2014.png

October 15, 2014

Sample 10A sequenced perfectly. Read to be transformed into yeast!

  • Need to submit FAM20 Kex + clone 9B for this full-coverage sequencing.