Technical data

5 Indexing and Sample Processing for Multiplexed Sequencing

Step 1A. Amplify the captured libraries with indexing primers containing 8-bp indexes A01–H12

70 SureSelect

Target Enrichment System for Illumina Multiplexed Sequencing

Step 1A. Amplify the captured libraries with indexing primers

containing 8-bp indexes A01–H12

In this step, the SureSelect-enriched DNA libraries are PCR amplified in

PCR reactions that include the appropriate indexing primer for each

sample.

This step uses the components listed in Table 37. Thaw then vortex to mix

the reagents listed below and keep on ice.

CAUTION

This chapter includes instructions for kits containing two different sets of indexing

primers. Verify that you are referencing the information appropriate for your kit

version before you proceed.

Step 1A covers kits with 8-bp indexes supplied in Library Prep Kit p/n 5500-0132 or

5500-0133 in format of white-capped tubes or blue plate (typically received December,

2014 or later).

Step 1B covers kits with 6-bp indexes supplied in Library Prep Kit p/n 5500-0105 or

5500-0075 in format of clear-capped tubes (typically received before December, 2014).

Table 37 Reagents for post-capture indexing by PCR amplification

Kit Component Storage Location

5× Herculase II Reaction Buffer Herculase II Fusion DNA Polymerase kit*, –20°C

100 mM dNTP Mix (25 mM each dNTP) Herculase II Fusion DNA Polymerase kit

, –20°C

* Do not use the PCR Reaction Buffer or dNTP mix from any other kit.

Herculase II Fusion DNA Polymerase Herculase II Fusion DNA Polymerase kit, –20°C

SureSelect ILM Indexing Post-Capture Forward

PCR Primer

SureSelect Target Enrichment Kit ILM Indexing

Hyb Module Box 2, –20°C

SureSelect 8 bp Indexes (reverse primers) SureSelect XT Library Prep Kit ILM, –20°C