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
increase dimer primers
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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