Brilliant II SYBR Green qPCR Master Mix

PCR Quantitative real-time PCR - Mammalian DNA

Experiment
PCR Quantitative real-time PCR - Mammalian DNA
Product
Brilliant II SYBR Green qPCR Master Mix from Integrated Sciences
Manufacturer
Integrated Sciences

Protocol tips

Upstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp.

Publication protocol

4.2. Selection of Primer Pairs for qPCR
Primers for qPCR were designed by Primer Express Software (Applied Biosystems, Foster City, CA, USA). PCR reactions (final volume 13 µL) contained 2× SYBR Green Master Mix (Brilliant II SYBR Green QPCR Master Mix, Agilent, Santa Clara, CA, USA), 2.7 ng of human cDNA or appropriate amount of the external standards (see below) and 2.5 pmol of primers for YKL-40. The PCR reactions were performed using Mx3005P QPCR System (Agilent) as follows: 10 min denaturation at 95 °C, 40 cycles of denaturation at 95 °C for 30 s, annealing at 55 °C for 1 min and polymerization at 72 °C for 1 min. Melting curves were generated after amplification. The PCR products were electrophoresed on a 10% polyacrylamide gel and analyzed using the Luminescent Image Analyzer (ImageQuant LAS 4000, GE Healthcare, Little Chalfont, UK). The primers’ sequences are listed in Figure S1. Chit1, AMCase, pepsinogen C, GAPDH and β-actin primers have been previously reported [45,46,47].



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Discussion

Discussion

5 years ago

Author: Germany

What is the optimal concentration for primers in qPCR?

What is the optimal concentration for primers in qPCR? My total volume is 20μl per reaction.

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing PCR Quantitative real-time PCR - Mammalian DNA using Brilliant II SYBR Green qPCR Master Mix from Integrated Sciences.

Paper title
Quantitative Real-Time PCR Analysis of YKL-40 and Its Comparison with Mammalian Chitinase mRNAs in Normal Human Tissues Using a Single Standard DNA
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Manufacturer protocol

Download the product protocol from Integrated Sciences for Brilliant II SYBR Green qPCR Master Mix below.

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