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Matermix / 一步法RT-qPCR试剂盒(染料法)ADVANCEDtm qPCR 1 step mastermix withSUPERGREENtm dye
wisent维森特
100 reactions
Ordering Information:
Catalogue number |
Quantity |
Specification |
801-001-AR |
100 reactions |
LOW ROX |
801-001-CR |
500 reactions |
LOW ROX |
801-001-LR |
1200 reactions |
LOW ROX |
801-002-AR |
100 reactions |
HIGH ROX |
801-002-CR |
500 reactions |
HIGH ROX |
801-002-LR |
1200 reactions |
HIGH ROX |
For instrument compatibility refer to APPENDIX I
Product Description:
WISENT ADVANCED 1-Step SUPERGREEN mix is used to quantify any RNA template including mRNA, total RNA and viral sequences. Our mix allows low copy number targets to be detected specifically with high efficiency. Proprietary small molecular inhibitor technology prevents formation of primer dimers leading to improved reaction sensitivity and specificity. Advanced buffer chemistry and polymerase technology offer market leading performance with minimal or no optimisation. Our mix is also ideal for fast and ultra fast cycling conditions.
Advanced technology for efficient cDNA synthesis and qPCR in a single tube features:
Thermostable reverse transcriptase 45°C to 55°C
Advanced RNase inhibitor
SUPERGREEN intercalating dye
Rapid extension rate for early Ct values
Best available sensitivity - increased limit of detection
Ideal on all real-time PCR platforms - standard and fast cycling conditions
Product Stability And Storage:
On arrival store at -20°C.
Avoid prolonged exposure to light.
If stored correctly the mastermix will retain full activity for 12 months. It is stable at 4°C for 1 month and can go through 30 freeze/thaw cycles with no loss of activity.
Instructions For Use:
Note: For efficient amplification under fast cycling conditions we recommend amplicon lengths between 80bp and 200bp. Do not exceed 400 bp. The shorter the amplicon length the faster the reaction can be cycled. Primers should have a predicted melting temperature of around 60°C, using default Primer 3 settings.
Reaction Setup:
1. Before starting, briefly vortex 2x WISENT ADVANCED 1 step mix .
2.Prepare a final mix , recommended 20 ul, as follows
Add |
Volume |
Final concentration |
WISENT ADVANCED 1 step mix |
10 ul |
1X |
Forward primer (10 uM) |
0.8 ul |
400 nM |
Reverse primer (10 uM) |
0.8 ul |
400 nM |
20 x RTase |
1 -2 ul |
1.0 ul is recommended . 2.0 ulmight |
Template RNA |
total RNA :1 pg to 1 ug mRNA : >0.01 pg |
variable |
Add PCR grade water up to 20 ul final volume |
3. Program your instrument using following conditions, acquiring data on the SYBR® Green or FAM channel:
Cycles |
Temperature |
Time |
Notes |
1 |
45 to 55 °C |
10 min |
45 °C:Recommended for most applications , 55 °C to be used only if amplicon contains region of high secondary structure |
|
95°C |
2 minutes |
Polymerase activation |
95°C |
5 seconds |
Denaturation |
|
60°C to 65°C |
20-30 seconds |
Anneal/Extension do not exceed 30 seconds ; do not use bellow 60 °C |
|
Melt analysis |
Refer to instrument instructions |
optional |
APPENDIX I
HIGH ROX / LOW ROX INSTRUMENT COMPATIBILITY
FOR FOLLOWING KITS:
-WISENT ADVANCEDtm qPCR MASTERMIX WITH SUPERGREENtm DYE
-WISENT ADVANCEDtm qPCR 1 step mastermix with SUPERGREENtm dye
-WISENT ADVANCEDtm qPCR PROBE 1 step mastermix
-WISENT ADVANCEDtm qPCR GENOTYPING mastermix
Manufacturer |
Instrument |
Lo-ROX |
Hi ROX |
Analytica Jena |
qTower |
Yes |
Yes |
Applied Biosystems |
7500, 7500 FAST, Viia7TM |
Yes |
No |
Applied Biosystems |
7000, 7300,7700,7900, 7900HT, 7900HT FAST, StepOneTM, StepOneTM Plus |
No |
Yes |
Bio-Rad® |
iCycler®, MyiQ®, iQ TM5, OpticonTM, OpticonTM2, Chromo4TM, MiniOpticonTM, CFX96TM, CFX384TM |
Yes |
No |
Cepheid® |
Smartcycler® |
Yes |
Yes |
Eppendorf |
Mastercycler® ep realplex, Mastercycler® realplex 2S |
Yes |
Yes |
Illumina® |
EcoTM |
Yes |
Yes |
Qiagen/Corbett |
Rotor-GeneTM 3000, 6000, Q |
Yes |
Yes |
Roche Applied Science |
Lightcycler®480, Lightcycler®Nano |
Yes |
Yes |
Stratagene (Agilent) |
MX 4000P®, MX 3000P®, MX 3005P® |
Yes |
No |
Takara |
Cycler Dice® |
Yes |
Yes |
Techne |
Quantica® |
Yes |
Yes |
Manufacturer |
Instrument |
Lo-ROX |
Hi ROX |
Note: when offered No-ROX is compatible with all machines above
热线:400-823-3718
网址:www.wisentbioproductschina.com
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