AlanFung:Protocol/Construction of Solexa sequencing Library: Difference between revisions
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*QPCR quantification | *QPCR quantification | ||
==Fragmentation and end-polishing== | ===Fragmentation and end-polishing=== | ||
*The PCR amplicons (200ng-1ug) are fragmented with Covaris sonicator to ~100bp | *The PCR amplicons (200ng-1ug) are fragmented with Covaris sonicator to ~100bp | ||
*End-Repair Reactions | *End-Repair Reactions |
Revision as of 17:58, 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 |
- Incubate tubes at RT for 30minutes.
- Perform a Qiaquick purification and elute with 39ul EB buffer.