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ADVANCEDtm qPCR PROBE 1 step mastermix

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ADCANCED™qPCR PROBE 1 step Mastermix
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1
介绍详情

 Ordering Information:

CAT.NO.

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

 

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Product Stability And Storage:

On arrival store at -25°C to -15℃.

Avoid prolonged exposure to light.

If stored correctly the mastermix will retain full activity for 12 months. It is stable at 2-8°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

Note: when offered No-ROX is compatible with all machines above. 

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