Calendar/2012-7-13

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Library Construction of Dr. Yi Zhang's DiP libraries Sample

Samples Preparation Protocol from Li

File:0712 DIP-seq samples.pdf

  • Received samples from Li on Jul 13 2012, 12 samples total in a yellow box, transfer to -20C

qPCR with N2 Barcodes

DIP Sample N2 Barcode
shCONT-input 1
shCONT-IgG 2
shCONT-5mC 3
shCONT-5hmC 4
shCONT-5fc 5
shCONT-5caC 6
shTDG-input 7
shTDG-IgG 8
shTDG-5mC 9
shTDG-5hmC 10
shTDG-5fC 11
shTDG-caC 12
Content Volume Per Reaction MM
Template 10 Do not add to MM
5uM PCR F+R Barcode 1 Do not add to MM
50X SYBG 0.2 2.64
2X PhusionHF MM 25 330
H20 13.8 182.16
Total Volume 50 660
  • Add 20ul 10uM N2-bar-code primers into strip tube
  • Add 20ul 10uM PCR_F primers and mix
  • We have 5uM F+R N2 Ind primer (Store at 4C Alice yellow 0.2ul tube rack)
  • Add 10ul template (for low volume sample add 5ul sample and add 5ul h2o) and 1ul barcode primers (using multi channel pipette) to strip tube
  • Add 39ul MM to each well
  98C 30sec 
  (98C 10sec -> 63C 20sec-> 72C 30sec) x 20 cycles
  72C 2min  
  15C hold.