One.Step RT-PCR Master Mix with SybrGreen, no-ROX, UNG/dUTP

250 rcs / 20μl2 x 1,25 ml

Mastermix for RT-real-time PCR based on a genetically engineered reverse transcriptase, contains SybrGreen as intercalating dye.

Features:
The kit contains all reagents required for RT-qPCR in a set to ensure fast and easy preparation with a minimum of pipetting steps. Just add template, primers. That's all.
     
The One.Step RT-PCR Kit is highly sensitive: less than 10 pg to 100 ng total RNA or < 1 pg to 20 ng poly(A) RNA (mRNA) can be detected when using highly expressed transcripts

Description:
The enzyme mix is based on a new formulated reverse transcriptase, a Hot-Start Polymerase and RNase Inhibitor. The set is providing increased specificity, high cDNA yield and improved efficiency for highly structured and long cDNA fragments. The reaction buffer includes extrapure dNTPs, SybrGreen as intercalating dye and reaction enhancers.

Principal:
In the RT-step the reverse transcriptase synthesizes single-stranded DNA molecules (cDNA) complementary to the RNA template. The hot-start polymerase activity is blocked.
In the first cycle of the PCR step the Hot-Start DNA polymerase activity is switched on and it synthesizes DNA molecules complementary to the cDNA, thus generating a double-stranded DNA template.

The One.step Realtime RT-PCR Master Mix from GeneON offers a tremendous convenience when applied to analysis of targets from multiple samples of RNA and minimizes the risk of contaminations.

Platforms: The Kit is suitable for all block-based Thermocycler. Stringent Quality Tests on ABI StepOne plus PCR Cycler


Sensitivity:
Targets can generally be detected from <1 pg to 20 ng poly(A) RNA (mRNA) or 10 pg to 1 μg total RNA. Even lower amounts of RNA may be successfully amplified by using highly expressed transcripts.

Components of Maximo.OneStep RT-qPCR:
Enzyme-mix: HotStart Taq Polymerase, Reverse Transcriptase, RNase Inhibitor and enhancers, Reaction buffer, extra pure dNTPs, SybrGreen, and stabilizers
RNase-free water

Optional: high ROX or UNG/dUTP (please refer to separate products numbers)

Storage:
@ -20°C, avoid frequent thawing and freezing, store all components with EvaGreen in the dark

Protocols / Manuals
The datasheet includes detailed protocols for:

1.) RT-PCR assay without sample denaturation (as a standard RNA/primer combinations)

2.) RT-PCR assay with sample denaturation (RNA/primer with a high degree of secondary
structure)
2.a) Preparation of RNA / Primer Mix
2.b) Denaturation and primer annealing
2.c) Preparation of the RT-PCR Mix

3.) Reverse Transcription and thermal cycling
Please see Product-Datasheet for all details below.