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.
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.
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
The absorption maximum of the inert dye is 615 nm.
@ -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.
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
- Add the volumes from the table to thin-
wall PCR tubes (preferred on ice)
|2x Re-action mix||12,5 µl|
|Bio-Star Mastermix||1,0 µl|
0,1 - 500 nM
0,1 - 500 nM
|RNA template||variable||1 pg - 1 µg|
|Sterile water||up to 25 µl|
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:
|1. Denaturation||92-94||3-5 min||1|
|Annealing||50-68 (Tm-5)||5-20 sec||25-45|
Bio-Star RT-qPCR SYBR Blue (2×) reagent kit includes 2× buffer for RT-qPCR containing all the necessary components (except for DNA template and primers), intercalating dye SYBR Green I, BioMaster-mix, DEPC-treated water and DMSO. The kit is designed for one-step reverse transcription and real-time polymerase chain reaction (RT-qPCR).
Final price excl. shipping costs3