One.Step RT-PCR Probe Kit with ROX

• 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 and the dual labeled fluorescent probe.
• 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; ROX and reaction enhancers. The basis is a Real-time PCR with dual labeled probes and multiplexing capacity.

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 kit is optimized for all real-time PCR cyclers who are compatible with the evaluation of the ROX reference signal.

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

Components of Maximo.OneStep RT-qPCR Master Mix (for probes):

Enzyme-mix: HotStart Taq Polymerase, Reverse Transcriptase, RNase Inhibitor and enhancers, Reaction buffer containing extra pure dNTPs and ROX
RNase-free water

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

Transport:
the product will be shipped with "blue ice"

For optimal results it is recommended to make an individually optimization for each RNA / primer pair.

Manufactured and quality-controlled in accordance with ISO 9001:2000

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)