One.Step RT-qPCR Kit (5X) "Perfect"  with separate ROX

Features:

The kit contains all reagents required for RT-qPCR (except template, primers and the dual labeled fluorescent probe) in one box to ensure fast and easy preparation with a minimum of pipetting steps. The premium quality enzyme mixes (5x PCR Mix and 20x RT Mix) and the optimized complete reaction buffer containing ultrapure dNTPs ensure superior real time PCR results. 

Description:

The enzyme mix is based on a with enhanced thermal stability providing increased specificity, high cDNA yield and improved efficiency for highly structured and long cDNA fragments.  

RT-qPCR is used to amplify double-stranded DNA from single-stranded RNA templates to allow a rapid real-time quantification of RNA targets. In the reverse transcription step the reverse transcriptase synthesizes single-stranded DNA molecules (cDNA) complementary to the RNA template. 

In the first cycle of the PCR step the hot-start DNA polymerase synthesizes DNA molecules complementary to the cDNA, thus generating a double-stranded DNA template. The hot-start polymerase activity is blocked at ambient temperature and switched on automatically at the onset of the initial denaturation. The thermal activation prevents the extension of non-specifically annealed primers and primer-dimer formations at low temperatures during PCR setup.

Advantages:

- High sensitivity

- Reverse transcription and real time PCR setup in one-step

- includes optimized RNase Inhibitor Reverse Transcriptase mixture

- Low risk of contamination and pipetting error

- Compatible with all standard real-time PCR cyclers

Platforms:

The Kit is suitable for all block-based Thermocycler.

-30 nM ROX: 

Applied Biosystems® 7500, 7500 Fast and ViiA™ 7, QuantStudio™ instruments, Agilent Mx3000P™, Mx3005P™, Mx4000™ and AriaMx.

-300 nM ROX: 

Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900 HT, StepOne™ and StepOnePlus™. 

Components  of Perfect- OneStep RT-qPCR Kit (5X):

Tube with 5X PCR complete-Mix;

Tube with 20X RT complete-Mix;

Tube with ROX 200µM for optional use;

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"

The kit and all components are produced in Europe

Protocols / Manuals: 

Considerations pre protocol:

Please note to protect RNA from degradation:

- prepare RNA samples in a separate laboratory area not used for setting  up reactions;

- use nuclease free gloves and labware;

- prevent the RNA degradation by spreading RNases;

- check the quality of the RNA sample on a denaturing agarose gel prior to cDNA synthesis;

The datasheet includes detailed protocol for:

Add the components to a nuclease-free micro-tube.

Pipette on ice and mix the components by pipetting gently up and down.

In general, water, RNA and primers should be mixed together before the rest of the components are added. 

Mix gently and run your RT-Cycler program.