Advanced PROBE 1 step Matermix / 一步法RT-qPCR试剂盒（探针法）价格

供应商：wisent维森特

规格：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

801-012-CR

500 reactions

801-012-LR

1200 reactions

801-013-AR

100 reactions

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 increase dimer primers
Probe(10μM)	0.4μL	200nM
Template RNA	total RNA: 1 pg to 1 μg mRNA : >0.01 pg	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