User guide

60
Fragment Library Preparation Using the AB Library Builder
™
System: 5500 Series SOLiD
™
Systems User Guide
AppendixC Supplemental Background Information
Preparing fragment libraries
C
Figure1 Fragment library preparation workflow overview
The barcoded adaptor consists of 3 segments of sequence:
• Internal adaptor sequence, which is necessary for sequencing the barcode
• Barcode sequence
• P2 Adaptor sequence, which is used for library amplification and emulsion PCR
Different libraries to be multiplexed in the same sequencing run are ligated to
barcoded adaptors with different barcode sequences. Ninety-six barcode sequences
are available to tag different libraries (see Figure 2).
Figure2 Fragment library design
Genomic DNA Sheared DNA
Library molecule ligated
with P1-T and barcoded
adaptors
P1-T Adaptor
Sheared DNA
P2 Adaptor (ss)
23 bp
T C T AC T T C T C C T T G G G C C C C G T C
P1-T Adaptor
Sheared DNA
5'
Barcoded Adaptor (ds)
19/53 bp
Barcode-0XX (ss)
10 bp
Internal Adaptor (ds)
20 bp
Internal Adaptor
P2 Adaptor
Barcode-
0XX
Internal Adaptor
C C CG
G G GC
T T
A A
G G
C C
C C
G G
G T
TC A
A C A
G
G C A G
C G T C
3'
T
T
Phosphorothioate bond
A T A G T G T T T T T T A T G G G A G T G G T G A T
T
CC C C C C C C C C C C C C C
CACCGACTGCAAG TAGAGGAAAGGG CAGGGGGG A T CT
P1-T Adaptor (ds)
41/42 bp
3'
5'
CC
CC
Barcoded Adaptor