User guide

29
Fragment Library Preparation Using the AB Library Builder
™
System: 5500 Series SOLiD
™
Systems User Guide
CHAPTER4
Nick-Translate the Library with
Optional Amplification
Workflow
Nick-translate the library
Prepare the
reaction, then nick-
translate the
library
Note: To nick-translate and amplify the library, proceed to “(Optional) Nick-translate
and amplify the library” on page 31.
1. In a new 1.5-mL LoBind Tube, combine for each library:
2. Vortex the reaction for 5 seconds, then pulse-spin.
3. Distribute 125-µL aliquots of combined library and PCR master mix between 4,
0.2-mL PCR tubes.
4. Incubate the library at 72°C for 20 minutes.
Nick-translate the libraries
(Optional) Nick-translate and amplify
the libraries
Prepare the reaction, then nick-translate
the library (page 29)
Purify the nick-translated library (page 30)
Prepare the reaction, then nick-translate
and amplify the library (page 31)
Purify the nick-translated, amplified
library (page 32)
Quantitate the DNA (page 33) Quantitate the DNA (page 33)
Stopping point Stopping point
Check the size distribution of the
library (page 34)
Check the size distribution of the
library (page 34)
Stopping point Stopping point
(Optional) Pool equal molar barcoded
libraries of similar size (page 34)
(Optional) Pool equal molar barcoded
libraries of similar size (page 34)
Stopping point Stopping point
Component Volume
Platinum
®
PCR Amplification Mix 400 µL
Library 100 µL
Total 500 µL