Technical data

SureSelect

Target Enrichment System for Illumina Paired-End

Sequencing Library Protocol

Agilent Technologies

Sample Preparation (200 ng DNA

Samples)

Step 1. Shear the DNA 42

Step 2. Assess quality (optional) 45

Step 3. Repair the ends 46

Step 4. Purify the sample using AMPure XP beads 47

Step 5. Adenylate the 3' end of the DNA fragments 49

Step 6. Purify the sample using AMPure XP beads 50

Step 7. Ligate the paired-end adaptor 51

Step 8. Purify the sample using AMPure XP beads 52

Step 9. Amplify the adaptor-ligated library 53

Step 10. Purify the amplified library with AMPure XP beads 56

Step 11. Assess quality and quantity 57

The sample preparation protocol is used to prepare DNA libraries for

sequencing using the Illumina paired-read platform. For each sample to be

sequenced, an individual indexed library is prepared. For an overview of

the SureSelect

target enrichment workflow, see Figure 1 on page 10.

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.

CAUTION

This section contains instructions for the preparation of gDNA libraries from 200 ng

DNA samples. For higher input (3

g) DNA samples, see the library preparation

protocol on page 21.