Technical data

6 SureSelect

Target Enrichment System for Illumina Multiplexed Sequencing

Contents

3 Sample Preparation (200 ng DNA Samples) 41

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

4 Hybridization and Capture 59

Step 1: Hybridize DNA samples to the Capture Library 60

Step 2. Prepare streptavidin-coated magnetic beads 66

Step 3. Capture the hybridized DNA using streptavidin-coated beads 67

5 Indexing and Sample Processing for Multiplexed Sequencing 69

Step 1A. Amplify the captured libraries with indexing primers containing 8-bp

indexes A01–H12 70

Step 1B. Amplify the captured libraries with indexing primers containing 6-bp

indexes 1–16 74

Step 2. Purify the amplified captured libraries using

AMPure XP beads 77

Step 3. Assess indexed library DNA quantity and quality 79

Step 4. Quantify each index-tagged library by QPCR (optional) 83

Step 5. Pool samples for multiplexed sequencing 84

Guidelines for sequencing sample preparation and run setup 86

6 Reference 87

Reference Information for Kits with Revised Index Configuration (indexing primers

in white-capped tubes or blue plate) 88