Overview

The TruSight HLA v2 Sequencing Panel protocol explains how to prepare indexed libraries from input DNA for subsequent cluster generation and DNA sequencing using the reagents provided in the Illumina TruSight HLA v2 Sequencing Panel. The goal of the protocol is to isolate and amplify HLA loci with a long-range polymerase chain reaction (PCR). After amplification, Nextera tagmentation fragments the DNA and leaves behind a sequence tag. An additional PCR then adds sequence adapters and indexing primers to generate DNA libraries ready for sequencing on the Illumina MiSeq systems.

The TruSight HLA v2 Sequencing Panel protocol provides:

• A single assay and workflow to obtain ultra-high resolution sequencing of 11 HLA Loci (Class I HLA-A, -B, -C; Class II HLA-DRB1/3/4/5, -DQA1, -DQB1, -DPA1, -DPB1)
• Unambiguous, phase-resolved HLA typing
• DNA-to-report solution includes an assay-optimized HLA analysis and reporting software, Assign 2.0 TruSight HLA Analysis Software
• Preparation of up to 24 samples per TruSight HLA v2 Sequencing Panel kit
• Gene coverage including exons and introns combined with paired-end sequencing allows for phase resolution across the entire gene, dramatically reducing the need to resolve phase ambiguities with follow-up assays

The protocol describes how to process 12 samples in parallel. The TruSight HLA v2 Sequencing Panel kit includes enough reagents to process 24 samples in total.

TruSight HLA v2 Sequencing Panel Protocol Guide (1000000010160 v00)