AmpliTaq Gold™ 360 DNA Polymerase

PCR Multiplex PCR - Bacterial DNA

Experiment
PCR Multiplex PCR - Bacterial DNA
Product
AmpliTaq Gold™ 360 DNA Polymerase from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

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

Multiplex PCR was performed in 15-μl reaction volumes consisting of 0.5 U of AmpliTaq Gold DNA polymerase (Applied Biosystems, Foster City, CA), 2 mM MgCl2, 250 μM each deoxynucleoside triphosphate (dNTP), 0.1 μM primer for khe-f and khe-r, 0.2 μM primer for ST258wzy-I-f and ST258wzy-I-r, and 0.4 μM primer for ST258wzy-II-f and ST258wzy-II-r in 1× PCR buffer (Applied Biosystems). The optimal cycling conditions were as follows: an initial denaturation step of 95°C for 10 min, followed by 35 cycles of 95°C for 30 s, 60°C for 30 s, 72°C for 1 min, and a final extension step of 72°C for 5 min. Amplicons were visualized after running at 120 V for 1.5 h on 1% agarose gels containing GelStar nucleic acid gel stain (Ionza Ltd., Allendale, NJ)

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Reviews

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Discussion

Discussion

5 years ago

Author: R. Verma India

Multiplex PCR and qPCR

I have been doing multiplex PCR and qPCR separately. Is there a way to combine them in a single reaction using Sybr Green?

Discussion

5 years ago

Author: Roland Lafontaine France

My multiplex PCR has decreased in sensitivity

Hello there! My multiplex PCR has decreased in sensitivity even though I have not changed any of the parameters. The materials are in good condition and the PCR machine is well calibrated. I have tested multiple samples but one of my targets has decreased in sensitivity.

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing PCR Multiplex PCR - Bacterial DNA using AmpliTaq Gold™ 360 DNA Polymerase from Thermo Fisher Scientific.

Paper title
Multiplex PCR for Identification of Two Capsular Types in Epidemic KPC-Producing Sequence Type 258 Strains
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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for AmpliTaq Gold™ 360 DNA Polymerase below.

Download PDF Download manufacturer protocol

Videos

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