Initially, these multiplex protocols were tested using different serial dilutions of genomic DNA extracted from peripheral blood (from 100 ng/mL to 5 pg/mL).
Thermal cycling profiles for the multiplex protocols were as follows: 15 min at 95°C, followed by 36 cycles of 30 s at 94°C, 90 s at 58 61°C and 60 s at 72°C, followed by final extension step of 10 min at 72°C.
Thermal cycling profiles for the multiplex protocols were as follows: 15 min at 95°C, followed by 36 cycles of 30 s at 94°C, 90 s at 58°C and 60 s at 72°C, followed by final extension step of 10 min at 72°C.
To best utilize the limited amounts of DNA obtained from microdissected specimens for the analysis of multiple loci, a two-step multiplex amplification protocol was adopted [ 23] to establish a 13-loci multiplex PCR (mPCR) (Additional file 3: Table S3).
A two-round multiplex PCR protocol was used to amplify these sequences from single sperm samples from nine unrelated healthy donors.
A similar multiplex PCR protocol was performed separately for detection of van genes by using specific primers for vanA, vanB, vanC-1, and vanC-2 genes (23).
A multiplex-PCR protocol was used to identify family-specific AmpC genes responsible for AmpC BL expression in organisms with or without a chromosomal AmpC BL gene [ 22].
For the detection of Brucella species, a multiplex real-time PCR protocol was used, targeting the bcsp31, alkB, and BMEI1162 genes of Brucella species, B. abortus, and B. melitensis, respectively, as previously described [ 17].
A novel multiplexed stripping voltammetric immunoassay protocol was designed for the simultaneous detection of multiple biomarkers (CA 125, CA 15-3, and CA 19-9 used as models) using PAMAM dendrimer-metal sulfide quantum dot (QD) nanolabels as distinguishable signal tags and trifunctionalized magnetic beads as an immunosensing probe.
Multiplex PCR protocols were designed for the simultaneous amplification of the QRDRs of topoisomerase genes (gyrA, parC and parE) and/or the genes involved in regulating the expression of the AcrAB efflux pump in their entirety (acrR, marRA operon and soxSR operon) (with a maximum of six PCR products).
