Technical data

5 Post-Capture Sample Processing for Multiplexed Sequencing
Step 4. Prepare samples for multiplexed sequencing
70 SureSelect
XT2
Target Enrichment System for Illumina
Step 4. Prepare samples for multiplexed sequencing
The final SureSelect
XT2
-enriched samples contain pools of either 8 or 16
indexed libraries, based on the Capture Library size and resulting
pre-capture pooling strategy.
The total number of indexed libraries that may be multiplexed in a single
sequencing lane is determined by the output specifications of the platform
used, together with the amount of sequencing data required for your
Capture Library. See Table 30 for sequence data requirement guidelines
based on Capture Library size. Calculate the number of indexes that can
be combined per lane, according to the capacity of your platform.
When appropriate for your sequencing platform, the 8-plex or 16-plex
samples may be further multiplexed by post-capture pooling, using the
guidelines provided in “Step 6. Optional: Pool captured libraries for
sequencing” on page 73. Prior to post-capture pooling, the DNA
Table 30 Sequencing data requirement guidelines
Capture Library Size Recommended Sequencing
Data per Indexed Library
Recommended Sequencing Data
per Pre-capture Pool
1 kb up to 499 kb 0.1 to 50 Mb* 1.6 to 800 Mb (16-index pools)
0.5 Mb up to 2.9 Mb 50 to 290 Mb
*
* For custom libraries, Agilent recommends analyzing 100X amount of sequencing data compared to
the Capture Library size for each sample. Pool samples according to your expected sequencing out-
put.
0.8 to 4.64 Gb (16-index pools)
3 Mb up to 5.9 Mb 300 to 590 Mb* 4.8 to 9.44 Gb (16-index pools)
6 Mb up to 11.9 Mb 600 to 1190 Mb* 9.6 to 19 Gb (16-index pools)
12 Mb up to 24 Mb 1.2 to 2.4 Gb* 19.2 to 38.4 Gb (16-index pools)
Human DNA Kinome 320 Mb 5.12 Gb (16-index pools)
Human All Exon v5 4 Gb 32 Gb (8-index pools)
Human All Exon v5 + UTRs 6 Gb 48 Gb (8-index pools)
Human All Exon v4 4 Gb 32 Gb (8-index pools)
Human All Exon v4 + UTRs 6 Gb 48 Gb (8-index pools)
Human All Exon 50 Mb 5 Gb 40 Gb (8-index pools)
Mouse All Exon 5 Gb 40 Gb (8-index pools)