User guide
34
Fragment Library Preparation Using the AB Library Builder
™
System: 5500 Series SOLiD
™
Systems User Guide
Chapter4 Nick-Translate the Library with Optional Amplification
Check the size distribution of the library
Check the size distribution of the library
Use 1 µL of sample in the Agilent Technologies 2100 Bioanalyzer
™
. If you see the
expected size distribution, proceed directly to emulsion PCR [refer to the SOLiD
™
EZ Bead
™
Emulsifier Getting Started Guide (Part no. 4441486)]. If you do not see the
expected size distribution, troubleshoot or contact your Life Technologies Applications
Specialist.
STOPPING POINT Store the DNA in Low TE Buffer at 4°C for short-term storage or at
–20°C for long-term storage; or proceed to “(Optional) Pool equal molar barcoded
libraries of similar size”.
(Optional ) Pool equal molar barcoded libraries of similar size
IMPORTANT! To avoid library bias, do not pool the libraries until after gel purification
if:
• the libraries are of dissimilar sizes
• it is unacceptable to pool libraries of unequal library representation
• you prefer not to pool libraries of similar sizes
1. Quantitate the libraries to be pooled by qPCR (see “Quantitate the DNA” on page
33.
2. Mix together equal molar amounts of each barcoded library of similar size in an
appropriately sized LoBind Tube. Vortex the tube.
3. (Optional) size-select the pooled libraries [see “(Optional) Size-select and pool
libraries” on page 51].
STOPPING POINT Store the library DNA in Elution Buffer (E1) at 4°C, or proceed directly
to templated bead preparation [refer to SOLiD
™
EZ Bead
™
Emulsifier Getting Started
Guide (Part no. 4441486)].