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ADVANCEDtm qPCR 1 step mastermix withSUPERGREENtm dye

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ADVANCED™Supergreen™qPCR 1 step Mastermix
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介绍详情

 Ordering Information:

CAT.NO.

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

 

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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: 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 

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