User-Supplied Consumables & Equipment

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Make sure that you have the required consumables and equipment before starting the protocol.

Some items are required only for specific workflows. These items are specified in separate tables.

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

Consumable

Supplier

1.7 ml microcentrifuge tubes

General lab supplier

10 µl pipette tips

General lab supplier

20 µl pipette tips

General lab supplier

200 µl pipette tips

General lab supplier

1000 µl pipette tips

General lab supplier

96-well 0.8 ml polypropylene deep-well storage plate (MIDI plate)

Thermo Fisher Scientific, part # AB-0859

Nuclease-free water

General lab supplier

Hard-Shell 96-well PCR plates

Bio-Rad, catalog # HSP-9601

Microseal 'B' adhesive seals

Bio-Rad, catalog # MSB-1001

Microseal 'F' foil seals

Bio-Rad, catalog # MSF-1001

RNase/DNase-free multichannel reagent reservoirs, disposable

VWR

[Optional] catalog # 89094-658

Ethanol 200 proof (absolute) for molecular biology (500 ml)

Sigma-Aldrich, product # E7023

Qubit dsDNA HS Assay Kit

One of the following, depending on quantification method:

• Thermo Fisher Scientific, catalog # Q32851
• Thermo Fisher Scientific, catalog # Q32854

Quant-iT PicoGreen dsDNA Assay Kit

Thermo Fisher Scientific, catalog # P11496

Qubit Assay Tubes

Thermo Fisher Scientific, catalog # Q32856

One of the following kits for quantification:

• Qubit dsDNA BR Assay Kit
• KAPA Library Quantification Kit – Illumina qPCR Instrument / Reference Dye Universal

 

• Thermo Fisher Scientific, catalog # Q32850 or Q32853
• Roche, catalog # 07960140001, KAPA code KK4824

One of the following kits, depending on qualification method:

Agilent Technologies, catalog #:

[Fragment Analyzer] High Sensitivity NGS Fragment Analysis Kit

DNF-474-0500

[Bioanalyzer] Agilent High Sensitivity DNA Kit

5067–4626*

[Dried Blood]Dried Blood Spots (DBS) card

GE Healthcare, catalog # 10534320

[Blood] EDTA blood collection tubes

Becton Dickinson

[Saliva] Oragene DNA collection kit for saliva

Genotek, catalog # OGR-500 or OGD-510

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

Consumables for Blood, Saliva, and Dried Blood Spot Input

Consumable

Supplier

[Dried Blood]Dried Blood Spots (DBS) card

GE Healthcare, catalog # 10534320

[Blood] Flex Lysis Reagent Kit

Illumina, catalog # 20018706

[Blood] EDTA blood collection tubes

Becton Dickinson

[Saliva] Oragene DNA collection kit for saliva

Genotek, catalog # OGR-500 or OGD-510

Equipment

Supplier

10 µl multichannel pipette

General lab supplier

10 µl single channel pipette

General lab supplier

20 µl multichannel pipette

General lab supplier

20 µl single channel pipette

General lab supplier

200 µl multichannel pipette

General lab supplier

200 µl single channel pipette

General lab supplier

1000 µl single channel pipette

General lab supplier

Magnetic Stand-96

Thermo Fisher Scientific, catalog # AM 10027

Microplate centrifuge

General lab supplier

Microcentrifuge

General lab supplier

Qubit Fluorometer 4.0

Thermo Fisher Scientific, catalog # Q33238 or Q33327

Vortexer

General lab supplier

One of the following analyzers:

• Fragment Analyzer
• 2100 Bioanalyzer Desktop System
• 4150 TapeStation System
• 4200 TapeStation System

Agilent Technologies catalog #:

• Refer to web product pages for catalog #
• G2939BA*
• G2992AA
• G2991BA

[Saliva] Water bath or air incubator capable of reaching 50°C

As recommended by DNA Genotek, refer to product pages.

* No longer available for purchase.

Thermal Cyclers

The following table lists the recommended settings for the thermal cycler. 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, separate

Bio-Rad part # 1851197¹

Bio-Rad DNA Engine Tetrad 2

Bio-Rad part # PTC-0240G²

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

MJ Research DNA Engine Tetrad

MJ Research part # MJ-T2-4

Eppendorf Mastercycler Pro S (Gradient S, Simulated Tube)

Eppendorf part # 41102404

¹ End of life announced. Refer to vendor site for more information.

² No longer available for purchase.