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Table 3 Categories classified by amplification specificity

From: The source of SYBR green master mix determines outcome of nucleic acid amplification reactions

Cat

ABI

Bio-Rad

Roche

Transcripts

Number

1a

Y

Y

Y

GAPDH, CD23, CD68, TLR9, Arg1, PDL1, CXCR4, COX2, B-actin, CXCR1, CCL2, CCL3, CD115, CD117, CD11b, CD163, CD14, CD66b, CD86, HLA-DR, IL10, HO-1, IL1b, IL6, S100A9, STAT4, STAT6, STAT3, TGFB1, TNFa, CCL5, CCL7, V-JUN, CSF3R-2, CD13-2, CCR5, CD31, CD44, CD54, CD64, CD16a, CD205, NFkB, S100A8, CCR2, CD62L, MSR1, CCL24, CD15, CD209, CLEC4C, FLT3, IFNγ

66

1b

Y

Y

Y/Na

IL-1RN, IL-18, CEBPB, v-FOS, Egr1, Egr2, CD54, CD200R, CD40, CD1c, TLR2, CD43, CCR7

2

Y

Y

N

CCL4

1

3

Neg

Y

Y

IL8, IL4

2

4

Neg

N

Y

CCL18

1

  1. Y a single smooth sharp peak; N more than two or unsmooth peaks; Neg no amplification
  2. aWith Roche mix, the primers generated specific PCR amplicons in the melting curve analysis, only when the suggested PCR protocol from the vendor’s guideline (Table 2) was used. With a general lab PCR protocol (Table 2) suboptimal melting curves were observed indicating additional aspecific PCR products