Add The Diluted Libraries To The Ion Chef; Library Sample Tube - Thermo Scientific Ion 520 User Manual

Chapter 4 Start the Ion Chef
4

Add the diluted libraries to the Ion Chef

2. Remove all cartridges and consumables from their packaging, then place them on the bench next
to the Ion Chef
Prepare the following cartridges and consumables:
• Chip Adapter and Chip Balance
• Enrichment Cartridge v2
™
• Ion S5
• PCR Plate and Frame Seal v2
• Recovery Station Disposable Lid v2 (2)
• Recovery Tube (12)
™
• Ion S5
™
• Ion S5
IMPORTANT! Before use, gently tap the Reagents and Solutions cartridges on the bench to force
the reagents to the bottoms of the tubes.
Note: When stored under normal conditions, a precipitate can form in some tubes of the Ion S5
ExT Chef Reagents cartridge. If present, load the cartridge as directed. The precipitate dissolves
when the reagents are mixed during instrument operation.

Add the diluted libraries to the Ion Chef

Tube
1. Pipet 50 µL of the diluted library, or combined library (see "Dilute the sample library"
into the Ion Chef
IMPORTANT! If you are performing a de novo sequencing experiment that does not include a
reference BAM file, add 4 μL of the Ion S5
™
Ion Chef Library Sample Tube. Cap the tube, vortex to mix, then centrifuge for 1–2 seconds to
collect contents at the bottom of the tube.
Note: If running the E. coli DH10B Control 400 Library, or the E. coli DH10B Control 600 Library,
dilute the stock library with nuclease-free water following the recommendations in "Dilute the
control library"
Tube.
2. Cap and store the diluted DNA library on ice, or at 4°C, until you are ready to load the tube into the
Reagents cartridge and the Ion Chef
26
™ run
™

Library Sample Tube

™
Instrument.
ExT Tip Cartridge
Chef Solutions cartridge
ExT Chef Reagents cartridge (from step 1)
™
Library Sample Tube (barcoded tube) from Position A of the Reagents cartridge.
on page 25, then pipet 50 µL of the diluted library into the Ion Chef
™
™
™
ExT Calibration Standard to the diluted library in the
Instrument.
Ion 520
Library Sample
on page
™
Library Sample
™ & Ion 530 ™ ExT Kit − Chef User Guide
™
25)