GSM5242317: SARS-CoV-2 titration - 80 - rep2; Homo sapiens; OTHER
We prepared 100x TCEP/EDTA buffer (250 mM TCEP, 100 mM EDTA, 1.15 N NaOH) 29. TCEP/EDTA buffer was added to human saliva at 1:100 volume, then samples were capped, vortexed to mix and heated in a thermocycler (95ºC 5 min, 4ºC hold) until ready to use for RT-LAMP. 1 μL of diluted LAMP material was used as a template for PCR using OneTaq DNA polymerase (NEB Cat. M0480L) with 100 nM each of custom dual-indexed Illumina P5 and P7 primers in either 10 or 25 μL reaction. PCR reactions were incubated as follows: (25 cycles of stage 1 [94ºC x 15 sec, 45ºC x 15 sec, 68ºC x 10 sec], 10 cycles of Stage 2 [ 94ºC x 15 sec, 68ºC x 10 sec], 68ºC x 1 min, 4ºC  x ∞).
GEO Accession
GSM5242317