AlanFung:LabNotes/CTCF/2010-6-7: Difference between revisions

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  Purification using qiaquick column
  Purification using qiaquick column
*Elute with 50ul buffer
*Elute with 50ul buffer
*Nanodrop result - 6.5ng/ul*50ul
 
Perform A-tailing protocol
*Add in 6ul dA-tailing Reaction buffer
*Add in 4ul klenow fragment
*Incubate at 37C for 30mins
 
 
Purification using qiaquick column
*Elute with 50ul buffer

Revision as of 16:43, 8 June 2010

Prepare PCR Product from Jurkat gDNA

ul
primer 1
h2o 7
template 2
Econo 2x 10
  • Program: 94C 2min-> (94C 40sec-> 52C 40sec -> 72C 60 sec) x 2 -> 72C 10min -> 15C hold
  • Qiaquick Purification
  • Nanodrop

P7-6.4ng/ul P10-5.4ng/ul

  • Gel Quant

File:ZhangLab 2 2010-06-07 14hr 34min.jpg

Protocol

Obtain 100ng Jurkat DNA
  • 100ng/(6.4ng/ul)=15.63ul
  • 100ng/(5.4ng/ul)=18.52ul
P7 P10
Jurkat DNA PCR Product 15.63 18.52
ddH2O 69.37 66.48
End Repair Reaction Buffer (10x) 10 10
Repair Enzyme Mix 5 5
  • Incubate 30 mins at 20C
Purification using qiaquick column
  • Elute with 50ul buffer
Perform A-tailing protocol
  • Add in 6ul dA-tailing Reaction buffer
  • Add in 4ul klenow fragment
  • Incubate at 37C for 30mins


Purification using qiaquick column
  • Elute with 50ul buffer