Delisted / NOT AVAILABLE -  Bio-Star one.step RT-PCR Master Mix with SybrGreen and blue inert Dye (2X)

Features/Description:

The one-step-kit contains all reagents required for RT-qPCR in a bundle to ensure fast and easy preparation with a minimum of pipetting steps. Just add template, primers. That's all.

• The mix is coloured for easy pipetting
• High specificity
• High sensitivity (1pg – 1 µg of RNA)
• Contains an inhibitor of RNases
• error-correcting activity because of Pfu DNA Polymerase
• recommended for GC-rich templates
• templates up to 5 kb

Description:

Bio-Star Master-mix contains the optimal ratio of M-MuLV–RH to HS-Taq DNA polymerase for both reactions.

M-MuLV–RH is a genetically modified Moloney Murine Leukemia Virus reverse transcriptase (M-MuLV). The enzyme has RNA- and DNA-dependent polymerase activity but lacks RNase H activity.

M-MuLV–RH reverse transcriptase exhibits improved thermal stability and is active at high temperatures (up to 50°С).

HS-Taq DNA polymerase is a recombinant Taq DNA polymerase inactivated by specific monoclonal antibodies.

The enzyme is inactive at temperatures up to 70 °C, it is activated at the first PCR cycle during a short 5-min incubation at 95 °C. Recombinant HS-Taq DNA polymerase catalyses 5’→3’ synthesis of DNA and possesses 5’→3’ exonuclease activity of the native Taq DNA polymerase from Thermus aquaticus.

Recombinant HS-Taq DNA polymerase is ideal for conventional PCR of the templates of up to 5 kbp.

Applications:

• Gene expression analysis;
• One-step real-time RT-qPCR.

Components RT-qPCR:

1. BioStar Enzyme-mix:

M-MuLV–RH reverse transcriptase, mixture of HS-Taq DNA Polymerase (inhibited with mAB), Pfu DNA Polymerase and inhibitor of RNases, stabilizer and enhancer

2.) Re-Action buffer (2X):  

100 mM Tris-НCl (рН 8.3 at 25 °С), 150 mM KCl, 0.6 mM each deoxynucleoside triphosphate, 6 mM MgCl2, 8 mM DTT, fluorescent dye SYBR Green I and inert dye, stabilizer

Note:

The absorption maximum of the inert dye is 615 nm.

Storage:

@ -20°C, short term at room temperature up to 2-3 days.

@ +4°C for about one week.

About 30 times Multiple thawing/freezing without any loss of activity.

Transport:

the product will be shipped with "blue ice"; up to 2-3 days @ room temperature.

Protocols / Manuals
The datasheet includes detailed protocols for your assay.

- Defrost and mix gently the 2x Re-action
  buffer

- Add the volumes from the table to thin-
  wall PCR tubes (preferred on ice)

Note: 

1.) In case of amplification of templates
     with complicated spatial structure,
     DMSO can be added in the amount of 1
     to 5% of the final volume of the reaction
     solution. Change in Тm of the primers
     should be taken into account when
     selecting the amplification program.

2.) Bio-Star qPCR Mastermix can volume
     can be in a range from 1-3 µl.

- Centrifuge your reaction chambers

- Start your PCR-Cycler Program following
  the table below: