Technical data

SureSelect

XT2

Target Enrichment System for Illumina Multiplexed

Sequencing Protocol

Agilent Technologies

Sample Preparation (100 ng DNA

Samples)

Step 1. Shear DNA 38

Step 2. Assess quality (optional) 41

Step 3. Repair the ends 43

Step 4. Purify the sample using AMPure XP beads 45

Step 5. Adenylate the 3' end of the DNA fragments 47

Step 6. Ligate the pre-capture indexing adaptor 48

Step 7. Purify the indexed DNA using AMPure XP beads 49

Step 8. Amplify the indexed library 50

Step 9. Purify the amplified library with AMPure XP beads 51

Step 10. Assess quality with the 2100 Bioanalyzer DNA 1000 Assay 52

This section contains instructions for the preparation of indexed gDNA

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

see the library preparation protocol on page 19.

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.