BioMaster RT-PCR – Color (2×)
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Application
Gene expression analysis;
One-step conventional RT-PCR.
RMС02-40
Number of reactions (50 μl)
40
€57
RMС02-200
Number of reactions (50 μl)
200
€250
RMС02-200
Number of reactions (50 μl)
200
€2,000
Basic description
BioMaster RT-PCR – Color (2×) includes 2× buffer for RT-PCR-Color containing all the necessary components (except for DNA template and primers), BioMaster-mix and DEPC-treated water. The kit is designed for one-step reverse transcription and polymerase chain reaction (RT-PCR).
BioMaster-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 exerts 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 catalyzes 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.
The buffer is optimized for both efficient RT and PCR. High density of the solution and marker dyes facilitate gel loading.
2× buffer for RT-PCR-Color
BioMaster-mix
DEPC-treated water
Number of reactions (50 μl each)
RMС02-40
2 × 0.5 ml
1 × 80 µl
0.5 ml
2 × 0.5 ml
40
RMС02-200
4 × 1.25 ml
2 × 200 µl
0.5 ml
3 × 1.8 ml
200
Product composition 2× buffer for RT-PCR-Color contains: 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, enzyme stabilizers and enhancers, marker dyes.
BioMaster-mix contains:
50 mM Tris-НCl (рН 8.0 at 25 °С), 100 mM NaCl, 1 mM EDTA, 5 mM DTT, 50 % (v/v) glycerol and 0.1 % (v/v) NP-40, M-MuLV –RH reverse transcriptase and HS-Taq DNA polymerase.
Limits of use
Not recommended for amplicons of ˃ 5 kbp.
Areas of use
Gene Expression Analysis RT-PCR one- and two-step Reaction mixture properties
The reaction mix is optimized for the specific and effective performance of M-MuLV –RH reverse transcriptase and HS-Taq DNA polymerase;
Allows long-term storage (storage of BioMaster RT-PCR – Color (2×) at room temperature for 2 days and/or multiple thawing-freezing cycles do not affect RT-PCR efficacy);
The mix contains dyes that do not interfere with polymerase performance and components increasing sample density for easy gel loading.
Note: Mobility of dyes in 0.5 – 1.5% agarose gel:
Xylene cyanol
Bromophenol blue
Orange G
Tartrazine
10000 – 4000 bp
500-400 bp
<100 bp
<20 bp
Benefits of using
High specificity
High sensitivity
Easy and convenient in use
Low pipetting error and low risk of cross-contamination
Standardized conditions of the same-type reactions (reduced pipetting error during mixing PCR components in a series of experiments)
PCR products can be further subjected to TA cloning due to deoxyadenosine overhangs at the ends of amplified DNA fragments.
Shorter step of sample preparation for the analysis of PCR results. No loading buffer is required due to the high density of the mixture.
Storage
9 months (under proper storage and transportation conditions).
Transportation
at -20 °С; not more than 30 thawing-freezing cycles.
Product passport
We carefully prepare our products and are ready to confirm their suitability with a passport.
Each box of our products has a unique lot number in the AA-000-0000 format.
