User-Supplied Consumables and Equipment

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The protocol has been optimized and validated using the items listed. Comparable performance is not guaranteed when using alternate consumables and equipment.

Consumables

Consumable

Supplier

Disposable gloves, powder free

General lab supplier

Eppendorf twin.tec PCR Plate 96, semiskirted, 250 µl, PCR clean

Eppendorf, catalog # 951020303

Ethyl alcohol, pure (500 ml)

Sigma-Aldrich, catalog # E7023

One of the following depending on assay throughput*:

Illumina, catalog #:
• Illumina Purification Beads, 30 ml
• 20119944
• Illumina Purification Beads, 100 ml
• 20060057
• Illumina Purification Beads, 400 ml
• 20060058

Lab tape

General lab supplier

Microseal 'B' PCR Plate Sealing Film

Bio-Rad, catalog # MSB1001

Microcentrifuge tubes, 1.5 ml

General lab supplier

Microcentrifuge tubes, 1.7 ml

General lab supplier

Microcentrifuge tubes, RNase-free, 1.7 ml

General lab supplier

Pipette tips, 10 µl

General lab supplier

Pipette tips, 20 µl

General lab supplier

Pipette tips, 200 µl

General lab supplier

Pipette tips, 1000 µl

General lab supplier

One of the following kits, depending on quality analysis method:

Agilent Technologies, catalog #:
• [Bioanalyzer System] Agilent DNA 1000 Kit
• 5067–1504
• [TapeStation System] D1000 ScreenTape
• 5067–5582
• [TapeStation System] High Sensitivity D1000 ScreenTape
• 5067–5584

Qubit Assay Tubes

Thermo Fisher Scientific, catalog # Q32856

Qubit dsDNA BR Assay Kit

Thermo Fisher Scientific, catalog # Q32850 or Q32853

RNase/DNase-free multichannel reagent reservoirs, disposable

VWR, catalog # 89094-658

Ultrapure water, nuclease-free

General lab supplier

* Each sample prepared with the Illumina RNA Prep with Enrichment, (L) Tagmentation protocol requires a total of 261 µl IPB. Preparation of 16 samples requires 4.2 ml IPB. Preparation of 96 samples requires 25.1 ml IPB. Illumina recommends purchasing 30 ml IPB for 96 samples.

Equipment

Supplier/Description

Multichannel pipettes, 10 µl

General lab supplier

Multichannel pipettes, 20 µl

General lab supplier

Multichannel pipettes, 200 µl

General lab supplier

Single channel pipettes, 10 µl

General lab supplier

Single channel pipettes, 20 µl

General lab supplier

Single channel pipettes, 200µl

General lab supplier

Single channel pipettes, 1000 µl

General lab supplier

Adhesive seal roller

General lab supplier

Magnetic Stand-96

Thermo Fisher Scientific, catalog # AM10027

Microplate centrifuge

General lab supplier

Microsample incubator

General lab supplier

Microsample incubator block, 1.5 ml

General lab supplier

Quality analysis instrument, any of the following:

Agilent Technologies, catalog #:
• 2100 Bioanalyzer System*
• G2939BA*
• 4150 TapeStation System
• G2992AA
• 4200 TapeStation System
• G2991BA

Thermal mixer, any of the following:

• BioShake iQ High-Speed Thermal Mixer
• BioShake XP High-Speed Thermal Mixer

Q Instruments, catalog #:

• 1808–0506
• 1808–0505

Vortexer

General lab supplier

[Optional] Amicon Ultra-0.5 centrifugal filter unit (0.5 ml, 30 kDa)

Millipore catalog # UFC503008

* End of life announced. Refer to vendor site for more information.

The following table lists the recommended thermal cyclers. If your lab has a thermal cycler that is not listed, validate the thermal cycler before performing the protocol.

Thermal Cycler

Supplier/Description

C1000 Touch Thermal Cycler with 96–Deep Well Reaction Module, bundled*

Bio-Rad part # 1840197*

C1000 Touch Thermal Cycler with 96–Deep Well Reaction Module, separate*

Bio-Rad part # 1851197*

T100 Thermal Cycler

Bio-Rad part # 1861096

Thermal cycler with the following specifications:

• Compatible with semi-skirted 96-well PCR plates
• Heated lid
• Average ramp rate: 2°C/sec to -2.5°C/sec
• Maximum ramp rate: 2.5°C to 4°C
• Maximum reaction volume: 100 µl
• Minimum temperature control range: 4°C to 100°C
• Minimum temperature accuracy: ±0.5°C
• Minimum temperature uniformity: ±0.5°C

General lab supplier

* End of life announced. Refer to vendor site for more information.