Technical data

SureSelect

XT2

Target Enrichment System for Illumina Multiplexed

Sequencing Protocol

Agilent Technologies

Sample Preparation (1 µg DNA

Samples)

Step 1. Shear DNA 20

Step 2. Assess quality (optional) 23

Step 3. Repair the ends 25

Instructions for Library Prep Kit p/n 5500-0102 or 5500-0103 26

Step 5. Adenylate the 3' end of the DNA fragments 29

Step 6. Ligate the pre-capture indexing adaptor 30

Step 7. Purify the indexed DNA using AMPure XP beads 31

Step 8. Amplify the indexed library 32

Step 9. Purify the amplified library with AMPure XP beads 34

Step 10. Assess quality with the 2100 Bioanalyzer DNA 1000 Assay 35

This section contains instructions for the preparation of indexed gDNA

libraries from 1 g DNA samples. For lower input (100 ng) DNA samples,

see the library preparation protocol on page 37.

For each sample to be sequenced, an individual indexed library is

prepared. For an overview of the SureSelect

XT2

target enrichment

workflow, see Figure 1 on page 12.

The sample preparation protocol is used to prepare DNA libraries for

sequencing using the Illumina paired-read sequencing platform. The steps

in this section differ from the Illumina protocol in the use of the Covaris

system for gDNA shearing, smaller target shear size, elimination of size

selection by gel purification, implementation of AMPure XP beads for all

purification steps, and primers used for PCR. Refer to the Illumina

protocol Preparing Samples for Multiplexed Paired-End Sequencing

(p/n1005361) or the appropriate Illumina protocol for more information.