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

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Construction of Solexa sequencing library from padlock captured PCR products using NEBNext DNA Sample Prep Master Mix Set 1 Kit (E6040S/L)
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
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Fragmented DNA'''
| align="center" style="background:#f0f0f0;"|'''85ul'''
|-
| 10X End Repair Bufer||10ul
|-
| End Repair Enzyme Mix||5ul
|-
|
|}

Revision as of 17:55, 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