User guide
21
Fragment Library Preparation Using the AB Library Builder
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System: 5500 Series SOLiD
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Systems User Guide
Chapter2 Prepare to Build the Library
Set up the AB Library Builder
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System for size-selected or express fragment library preparation
4.
Load the reagent cartridges into the cartridge rack by sliding each reagent
cartridge along the groove in the direction of the arrow until the reagent cartridge
clicks into place. Make sure that the notches in the cartridge align with the
notches in the cartridge rack.
Note: An incorrectly loaded cartridge rack may cause the instrument to stop
during a run.
Load adaptors in the cartridges
IMPORTANT! If you are preparing barcoded libraries for multiplexed sequencing, for
each sequencing run, use at least one of the following full sets of four barcodes:
Barcodes-T-001–004, 005–008, 009–012, 013–016, 017–020, 021–024, 025–028, 029–032,
033–036, 037–040, 041–044, 045–048, 049–052, 053–056, 057–060, 061–064, 065–068, 069–
072, 073–076, 077–080, 081–084, 085–088, 089–092, or 093–096. Use only one of the
barcoded-T-0XX adaptors for each ligation reaction, unless < 4 libraries are being
barcoded.
Use the barcodes according to these conditions:
• If <4 libraries are prepared for sequencing, then use multiple barcodes per library in
equal ratios. For example, for 2 libraries, use 2 barcodes for each library. For
3 libraries, use 4 barcode adaptors for each library for a total of 12 barcodes.
•If ≥4 libraries are prepared for sequencing and libraries are split into sets of 4 to use
full sets of barcodes, then use one set of barcodes for the remaining libraries (1,2,or
3 libraries). There is no need to use multiple barcodes per library in equal ratios.
1. Prepare the 5✕ Reaction Buffer tubes:
a. Spin the supplied 5✕ Reaction Buffer tubes for each sample.