The reactions were conducted in singleplex with a standard super mix (Bio-Rad iQ™ Multiplex Powermix) using RT-PCR Chromo4 (Bio-Rad) and Opticon Monitor 3 Software.
First-strand cDNA was synthesized using a Trans Script First-Strand cDNA Synthesis Super Mix (TransGen Biotech, China), according to the manufacturer's protocol.
Triplicate quantitative PCR assays per sample were performed using a Platinum SYBR Green Super mix (Invitrogen) with ROX reference dye according to the manufacturer's protocol.
He will watch from Denver, scouring tapes and his notes on free agency, breaking down the draft, signing some veteran Broncos, releasing a few, searching for that super mix that can once again plant him in the super game.
Quantative PCR (qPCR) was performed using iQ SYBR Green Super Mix and a MyiQ Single-Color PCR thermal cycler (Biorad, Hercules, CA).
The assay was performed with TransScript II Green One-Step qRT-PCR Super Mix (TransGen) using a CFX96 Real-Time PCR Detection System (Bio-Rad).
About 10 mg lung or spleen tissues were homogenized in Trizol reagents (Invitrogen, Carlsbad, USA), total RNA was extracted, and cDNA was reversely transcribed using an EasyScript First-Strand cDNA Synthesis Super Mix (TransGen Biotech, China) according to the manufacturer's instruction.
qPCR was performed on a MX3000P machine (Stratagene) using SYBR GreenER qPCR super mix (Invitrogen).
qPCR assays were performed using an iCycler (BioRad Laboratories, USA) and the Sybergreen Super mix (BioRad Laboratories, USA).
Polymerase chain reaction (PCR) reactions were performed on a 96-well optical plate with the Platinum Super Mix (Invitrogen, Paisley, UK) using 5 μl of diluted cDNA (equivalent to 10 ng total RNA), 200 n M of the probe and 400 n M primers in a 25 μl final reaction mixture.
