AlanFung:Protocol/Construction of Solexa sequencing Library: Difference between revisions
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>Alan6017518 (New page: Construction of Solexa sequencing library from padlock captured PCR products using NEBNext DNA Sample Prep Master Mix Set 1 Kit (E6040S/L)) |
>Alan6017518 No edit summary |
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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 |