AlanFung:Protocol/Construction of Solexa sequencing Library: Difference between revisions

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>Alan6017518
>Alan6017518
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*End-Repair Reactions
*End-Repair Reactions
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|Fragmented DNA||85ul
| Fragmented DNA||85ul
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| 10X End Repair Bufer||10ul
| 10X End Repair Bufer||10ul

Revision as of 17:56, 29 October 2009

Construction of Solexa sequencing library from padlock captured PCR products using NEBNext DNA Sample Prep Master Mix Set 1 Kit (E6040S/L)

Overview

  • Fragmentation and end-polishing
  • Size Selection
  • Ligation
  • PCR of sequencing Library
  • QPCR quantification

==Fragmentation and end-polishing

  • The PCR amplicons (200ng-1ug) are fragmented with Covaris sonicator to ~100bp
  • End-Repair Reactions
Fragmented DNA 85ul
10X End Repair Bufer 10ul
End Repair Enzyme Mix 5ul