APHL COVIDSeq Training Checklist

Before We Start (Instruments/Consumables/Reagent/Cycle) Check

Reference: illumina-covidseq-test-reference-guide-ruo-1000000126053-04

Extract RNA

Consumables

ELB HT (Elution Buffer HT)

CPC HT (COVIDSeq Positive Control HT)

1.7 ml LoBind tubes u 5 ml LoBind tubes

[Quick-DNA/RNA Viral MagBead] 2000 µl 96 deep well plate

Anneal RNA

Consumables

EPH3 HT (Elution Prime Fragment 3HC Mix)

96-well PCR Plate

Microseal 'B' adhesive seals

Synthesize First Strand cDNA

Consumables

FSM HT (First Strand Mix HT)

RVT HT (Reverse Transcriptase HT)

1.7 ml tubes (1 per 96-well sample plate)

Microseal 'B' adhesive seal

Amplify cDNA

Consumables

IPM HT (Illumina PCR Mix HT)

CPP1 HT (COVIDSeq Primer Pool 1 HT)

CPP2 HT (COVIDSeq Primer Pool 2 HT)

Nuclease-free water

15 ml tube (2 for four 96-well sample plates)

96-well PCR plates (3)

Microseal 'B' adhesive seal

Tagment PCR Amplicons

Consumables

EBLTS HT (Enrichment BLT HT)

TB1 HT (Tagmentation Buffer 1 HT)

Nuclease-free water u 1.7 ml tube

15 ml tube (1 per four 96-well sample plates)

Microseal 'B' adhesive seal

Store EBLTS HT upright at temperatures above 2°C. Make sure beads are always submerged in the buffer.

Post Tagmentation Clean Up

Consumables

ST2 HT (Stop Tagment Buffer 2 HT)

TWB HT (Tagmentation Wash Buffer HT)

Microseal 'B' adhesive seal

Amplify Tagmented Amplicons

Consumables

EPM HT (Enhanced PCR Mix HT)

Index adapters (IDT for Illumina-PCR Indexes Set 1, 2, 3, 4)

Nuclease-free water

15 ml tubes (1 per two 96-well sample plates)

96-well PCR plate

Pool and Clean Up Libraries

Consumables

ITB (Illumina Tune Beads)

RSB HT (Resuspension Buffer HT)

Freshly prepared 80% ethanol (EtOH)

1.7 ml tube (2 per 96-well sample plate)

PCR 8-tube strip