Universal SYBR Green Mixes for RT-qPCR
The universal SYBR Green quantitative PCR mixes are made for use with all types of qPCR devices.
Mixes contain SYBR Green mix (Taq polymerase, dNTPs, MgCl2 and SYBR Green) and 2 different ROX dyes: High Rox and Low Rox. Simply add the corresponding dye for the qPCR device you want to use.
The advantage of the universal SYBR Green mixes is therefore to be able to use the same mix for different devices.
Here's how to select the appropriate ROX dye for the qPCR device you are using:
| Universal | All instruments |
| Without ROX | Bio-Rad
CFX96, CFX384, iCycler iQ, iQ5, MyiQ, MiniOpticon, Opticon®, Opticon 2,
Chromo4; Cepheid SmartCycler®; Eppendorf Mastercycler® ep realplex, realplex 2 s; Illumina Eco qPCR; Qiagen/Corbett Rotor-Gene® Q, Rotor-Gene® 3000, Rotor-Gene® 6000; Roche Applied Science LightCyclerTM 480; Thermo Scientific PikoReal Cycler. |
| Low ROX | Applied
Biosystems 7500, 7500 Fast, ViiA7; Stratagene MX4000, MX3005P, MX3000P |
| High ROX | Applied
Biosystems 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast; StepOne, StepOnePlus |
For RT-qPCR applications, there are two approaches: one-step RT-qPCR and two-step RT-qPCR. Each of these methods has its advantages and disadvantages. As its name implies, single-step RT-qPCR combines the synthesis of complementary DNA (cDNA) of the first strand (reverse transcript, RT) and subsequent quantitative PCR in a single reaction tube. The two-step RT-qPCR involves two distinct reactions, starting with first strand cDNA synthesis (reverse transcription-RT) and then amplifying part of the resulting cDNA by quantitative PCR in a separate tube .
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