Promega PCR Master Mix

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp	PCR plates, tubes and tips should be UV sterilized for 20-30 mins

Upstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp
Protocol tips
PCR plates, tubes and tips should be UV sterilized for 20-30 mins

Publication protocol

Polymerase Chain Reaction experiment
PCR was performed using the extracted DNA from each of the samples. A total reaction volume of 20µl was used, which contained 10µl Taq mix (Promega, USA), 1µl each of 10µM both forward and reverse primer solutions, 1mM MgCl2, 3µl RNase/ DNase free water and 5µl DNA template extracted from samples. A negative control was performed by replacing 5µl of DNA template with water whiles a positive control contained 5µl of DNA extracted from pure cultures of each organism. PCR assays were performed in Gene-Pro PCR machine (Alpha Laboratories, UK).

The PCR progamme used consisted of initial denaturation, 94°C for 5mins, 30 cycles for denaturation, 94°C for 1min, annealing, 45°C, 60°C and 55°C for E. coli, S. aureus and Salmonella sp. respectively for 30s, and extension, 72°C for 1min and a final extension at 72°C for 5mins. After PCR, 10µl of the PCR product was run on 1% gel. Bands were observed under UV light and photographs taken using Epichemi imager (UVP Bio imaging Systems, UK).


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Manufacturer protocol

Download the product protocol from Promega for Promega PCR Master Mix below.

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