Brilliant II SYBR Green qPCR 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.

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].



Full paper   Login or join for free to view the full paper.

Reviews

Discussion

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.

Manufacturer protocol

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

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing PCR Quantitative real-time PCR - Mammalian DNA using Brilliant II SYBR Green qPCR Master Mix from Integrated Sciences. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.