AlanFung:LabNotes/CTCF/2010-4-23: Difference between revisions

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(Created page with '==Project Update== ===Bottleneck=== *Extreme low amount of ligated product *Low adapter ligation efficiency ===Improvements=== *Start of with high amount of input DNA (500ng & …')
 
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===Improvements===
===Improvements===
*Start of with high amount of input DNA (500ng & 1ug)
*Start of with high amount of input DNA (500ng & 1ug,50ng)
*Increase the amount of adapter and increase annealing time
*Increase the amount of adapter and increase annealing time



Revision as of 17:24, 23 April 2010

Project Update

Bottleneck

  • Extreme low amount of ligated product
  • Low adapter ligation efficiency

Improvements

  • Start of with high amount of input DNA (500ng & 1ug,50ng)
  • Increase the amount of adapter and increase annealing time

Quantification of Jurkat PCR Product

Qubit: 48.4ng/ul

Library Prep with new Jurkat PCR product

  • Concentration:48.4ng/ul
  • BP:100BP
  • setup: 1ug, 500ng and -ve control

End Repair

1ug 500ng 50ng -ve
DNA 20.66 10.33 1.03 0
End Repair Buffer 10 10 10 10
End Repair Enzyme Mix 5 5 5 5
ddh2o 64.34 74.67 83.97 85
Total Volume 100 100 100 100
  • Incubate at 20C for 30 minutes
  • EtOH purification resolve with 42ul ddh2o (250ul 100%EtOH, 10ul 3M NaOH, 1ul Glycoblue)