illumina HiSeq 2000, Системное руководство, Страница: 33 / 76

Chapter 4 Sequencing in TruSeq v3 Mode
Sequencing in TruSeq v3 Mode
Introduction 26
Prepare Reagents for Read 1 27
Enter Run Parameters 30
Load and Prime Reagents 33
Load the Sequencing Flow Cell 37
Monitor the Run 39
Prepare Reagents for Read 2 40
Unload Reagents 43
Perform a Water Wash 43
Introduction
To perform a sequencing run in TruSeq v3 mode on the HiSeq 2000, prepare SBS reagents for Read 1 and
indexing reagents before setting up the run. Follow the software prompts to set up the run. Run setup
includes entering run parameters, loading and priming reagents, loading the flow cell, and checking fluidics.
After the completion of Read 1 and any index reads, prepare and load paired-end reagents and SBS
reagents for Read 2.
Visit the HiSeq 2000 specifications page on the Illumina website for information about run duration and other
performance specifications.
Staggering Runs
You can start a new run on either flow cell A or flow cell B when a run on the adjacent flow cell is in progress.
For more information, see
Stagger Runs on Flow Cell A and Flow Cell B on page 52 .
Run Types for TruSeq v3 Chemistry
The following table shows types of sequencing runs and the number of possible cycles for each read when
using TruSeq v3 chemistry. Reference this information when setting up the run.
Run Type Read 1
Cycles Index 1
(i7) Read
Cycles Index 2
(i5) Read
Cycles Read 2
Cycles Total
Cycles
Single-Read,
Non-Indexed ≤ 101 -- -- -- ≤ 101
Single-Read,
Single-Indexed ≤ 101 6 or 7 ¹
8 ² -- -- ≤ 108 ¹
≤ 109 ²
Single-Read,
Dual-Indexed ≤ 101 8 8 -- ≤ 117
Paired-End,
Non-Indexed ≤ 101 -- -- ≤ 101 ≤ 202
Paired-End,
Single-Indexed ≤ 101 7 ¹
8 ² -- ≤ 101 ≤ 209 ¹
≤ 210 ²
Paired-End,
Dual-Indexed ≤ 101 8 7 + 8 ³ ≤ 101 ≤ 225
¹ Number of cycles for single-indexed libraries
² Number of cycles for dual-indexed libraries
³ The Index 2 Read of a paired-end dual-indexed run includes 7 additional chemistry-only cycles
Document # 15011190 v03
For Research Use Only. Not for use in diagnostic procedures.
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