Multiplex PCR 5X Master Mix

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Multiplex with flexibility and minimal optimization

  • Simultaneously detect two or more products in a single reaction
  • Enhanced buffer formulation for optimal performance 
  • 5X master mix format for easy reaction setup - just add template DNA and primers

Ordering Information

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  • 5 X
    100 reactions (50 μl vol)
    $228.00
  • Product Information

    Multiplex PCR can simultaneously detect two or more products in a single reaction. There is an increasing demand for multiplex PCR techniques in assays conducted in research laboratories and forensic/diagnostic genotyping assays (1,2). Multiplex PCR can also be used for semi-quantitative gene expression analysis using cDNA templates. The NEB Multiplex PCR 5X Master Mix is an easy-to-use solution featuring high quality recombinant Taq DNA Polymerase. The mix is optimized for high yield and robust performance. Its performance is illustrated in a 15-plex PCR using human genomic DNA (Figure 1) and an 8-plex PCR using cDNA products as templates (Figure 2). The 5X formulation allows the user maximal input of primers, template DNAs and additional components.

    Figure 1
    Figure 1:
    15-plex PCR using varying amounts of human genomic DNA. 1X Multiplex PCR 5X Master Mix was used with 0.15 μM of each primer. The cycling conditions were 95°C for 1 minute, 35 cycles of 95°C for 20 seconds, 60°C for 1 minute and 68°C for 2 minutes. Marker M is the 2-Log DNA Ladder (NEB #N3200).
    Figure 2
    Figure 2
    8-plex PCR using cDNA products from 1 ng human spleen total RNA. Cycling conditions were 95°C for 1 minute, 30 cycles of 95°C for 20 seconds, 60°C for 30 seconds and 68°C for 2 minutes. Amplicon lengths are indicated next to gel.
    Figure 3
    Figure 3:
    Comparison of PCR product yields between single-plex and multiplex PCR reactions. Figure A shows amplification of human genomic DNA fragments (sizes are listed to the left of the gel). Lane 1 is a 3-plex reaction, Lanes 2-4 are single-plex reactions, and Lanes 5–6 are 2-plex reactions; Lane 7 is the 1 kb DNA Ladder (NEB #N3232 ). Figure B shows the results of expression analysis of five mRNAs using cDNA templates (first strand synthesis was carried out using the ProtoScript Kit with human spleen total RNA); Lane 1 is a 5-plex PCR from cDNA reactions, while Lanes 2–6 are single-plex PCR reactions; Lane 7 is the 100 bp DNA Ladder (NEB #N3231).

    Product Source

    An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1.
    Product Categories:
    Taq DNA Polymerase Products,
    Master Mix Products
    Applications:
    Multiplex PCR,
    Specialty PCR
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