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Advanced PROBE
1 step Matermix / 一步法RT-qPCR试剂盒（探针法）wisent维森特
500 reactions
Ordering Information:
| Catalogue number |
Quantity |
Specification |
| 801-011-AR |
100 reactions |
LOW ROX LOW ROX LOW ROX |
| 801-011-CR |
500 reactions |
|
| 801-011-LR |
1200 reactions |
| 801-012-AR |
100 reactions |
HIGH ROX HIGH ROX HIGH ROX |
| 801-012-CR |
500 reactions |
|
| 801-012-LR |
1200 reactions |
| 801-013-AR |
100 reactions |
NO ROX NO ROX NO ROX |
| 801-013-CR |
500 reactions |
|
| 801-013-LR |
1200 reactions |
For instrument compatibility refer to APPENDIX I
Product Description:
WISENT ADVANCED PROBE 1-STEP mix uses advanced reverse transcriptase technology and buffer chemistry for efficient cDNA synthesis.
This kit features:
Thermostable and extremely active RTase.
RNAse inhibitor preventing degradation of RNA
Buffer system that allows efficient amplification from GC rich and AT rich templates
Real-time PCR probe mixes designed for use on a wide range of technologies including TAQMAN R , MOLECULAR BEACONS R , AND SCORPION PROBES R
Suitable for total RNA as a substrate - RTase is not inhibited by ribosomal and tRNA
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:
1 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.
2 For TAQMANR probes choose probe close to 5’ primer and avoid terminal guanosine residues.
Reaction Setup:
1 Before starting, briefly vortex 2x WISENT ADVANCED 1 step mix .
2 Prepare a final mix , recommended 20 μL, as follows:
| Add |
Volume |
Final concentration |
| WISENT ADVANCED 1 step mix |
10 μL |
1x |
| Forward primer (10 μM) |
0.8 μL |
400 nM |
| Reverse primer (10 μM) |
0.8 μL |
400 nM |
| 20 x RTase |
1 -2 μL |
1.0 μL is recommended . 2.0 μL might |
| Probe(10μM) |
0.4μL |
200nM |
| Template RNA |
total RNA: 1 pg to 1 μg |
variable |
| Add PCR grade water up to 20 μL 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 |
| 1 |
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 |
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 |
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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