SYBR Green PCR Premix HS Taq(Real Time)

Cat.No.: GR1201-1ML / GR1201-5ML

Size: 50μl×40T/50μl×200T

Trait: High efficiency, strong specificity, stable storage.amplification
Description
   The product is used in Real Time PCR by SYBR® Green I tabling fluorescence method. It is a 2×premix containing SYBR® Green I. Only template, primer and ddH2O needed to be added into PCR reaction system. It simplifies operation and reduces pollution during experiment. The product is easy to use and makes the experimental data reliable.
   The product contains newly developed Hot Start Taq DNA polymerase which can effectively inhibits nonspecific PCR amplification, greatly improves the efficiency of PCR amplification and enhances the sensitivity of the Real Time PCR.
Features
Used in Real Time PCR, can accurately detect and quantify the target gene.
2×Premix contains Hot Start DNA polymerase, dNTPs, SYBR Green I. Only template, primer and ddH2O should be added during PCR reaction. So it is very convenient to use.
DNA polymerase is an improved Hot Start DNA polymerase and buffer system is also optimized, both make the product has high amplification efficiency and strong specificity.
Usage: Used in Real Time PCR.
Storage: -20
Protect from light. Subpackage and store at -20℃ for long time. Store at 4℃ for 3 months.
Note:
This product is only used for scientific research, not applied to diagnosis or clinical trials. Please strictly abide by the laboratory safety operation during using.
Applicable instrument
ABI PRISM®7000, 7700 and 7900 series
LightCycle® series
Bio-Rad
Components
F   HS Taq DNA Polymerase
F   2 ×reaction buffer
F   3 mM MgCl2
F   0.4 mM dNTPs
F   2×SYBR Green
F   PCR improver
Principles for primer design
F   Length: 2030bp
F   GC%: 40%60%
Primer should be close complementary with temple
Avoid continuous same base at 3’ end.
Amplified fragments: ≤300bp (optimum for 100bp~300 bp )
 
Template
RNA template: The product is not suitable for one step RT-PCR, the RNA template must reverse transcribed to cDNA for PCR amplification. The template concentration can be low and the cDNA can be diluted before use. Suggest using first -strand cDNA synthesis kit (Cat.No: TER016-1 and TER016-2 )
DNA template: The product is ready for quantitative PCR amplification. The DNA template should be 1~10ng without protein or RNA pollution. DNA can be obtained by Genview nucleic acid purification system: High efficient centrifugal column DNA product purification kit (Cat.No: GV-PCR-P-50) or High efficient centrifugal column agarose gel DNA recovery kit (Cat.No: GV- GX-50).
 
ExampleABI PRISM®7700
PCR reaction system (take mouse tissue DNA as template, amplify 318bp gene fragment of β-actin in 25μl reaction system)

DNA template
110 ng
β-actin R(5μM)
0.5 μl
β-actin F(5μM)
0.5 μl
SYBR Green PCR Premix HS Taq
12.5 μl
H2ORNase free
to 25μl

 
PCR cycle
94   3 min
 
 
 
 
Notes
Annealing temperature is only for the example of primer and template. User should determine the annealing temperature based on actual Tm value of primer.
Fluorescence quantitative PCR instrument collect data at 72℃ extension step, the extension time should ≥30s using ABI PRISM® series.
Strictly operate by fluorescence quantitative PCR instrument software
Use import 0.2ml PCR tube.
 

 

 

 

 

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