AlanFung:LabNotes/CTCF/2010-4-23

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Revision as of 17:23, 23 April 2010 by >Alan6017518 (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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Project Update

Bottleneck

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

Improvements

  • Start of with high amount of input DNA (500ng & 1ug)
  • 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)