title: StepOnePlus Real-Time PCR System human PBMCs
status: Public on Jan 05 2022
contact.city: Rotterdam
contact.first: Aafke
contact.last: Duizendstra
contact.address: Doctor Molewaterplein 40
contact.country: Netherlands
contact.institute: Erasmus Medical Center
organism: Homo sapiens
description: PCR assay
manufacture_protocol: Forward and reverse primers were designed with NCBI PrimerBLAST according to MIQE guidelines. To prevent co-amplification of genomic DNA, intron-flanking primers or exon-exon junction primers were designed with target amplicon sequences of 80 to 150 bp with a maximum GC content of 65%. Forward and reverse primers were not modified and were purified with a desalt step (Merck, Darmstadt, Germany). Primer pairs were tested in duplicate for their optimal annealing temperature and amplification efficiency using healthy control PBMC derived cDNA.