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ADVANCEDtm qPCR GENOTYPING mastermix

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ADVANCED™qPCR GENOTYPING Mastermix
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介绍详情

Ordering Information:

CAT.NO.

Quantity

specification

801-020-AR

100 reactions

LOW ROX

LOW ROX

LOW ROX

801-020-ER

500 reactions

801-020-LR

1200 reactions

801-021-AR

100 reactions

HIGH ROX

HIGH ROX

HIGH ROX

801-021-ER

500 reactions

801-021-LR

1200 reactions

801-022-AR

100 reactions

NO ROX

NO ROX

NO ROX

801-022-ER

500 reactions

801-022-LR

1200 reactions

For instrument compatibility refer to APPENDIX I 

Product Description:

This kit features:

Accurate allelic discrimination

Superior allele clustering

Compatible with Taqman R dual-labeled probes

Compatible with all real-time PCR platforms

Our Genotyping Mix is a kit designed for use in dual-labeled probe based genotyping assays including Taqman™ genotyping assays. The mix is fully compatible with ABI TaqMan Pre-Designed SNP Genotyping assays.

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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. The probe Tm should be approximatelly 10 °C .Higher than the primer Tm.

3 Template: Use between 1 and 20 pg human genomic DNA per reaction. For accurate allele calling similar amounts of template must be used in all wells of the same run.

Reaction Setup:

1. Before starting, briefly vortex 2x WISENT genotyping mix

2. Prepare a final mix , recommended 20 ul, as follows:

Add:

Volume

Final concentration:

WISENT genotyping  mix

10 ul

1X

Forward primer (10 uM)

0.8 ul

400 nM

Reverse primer (10 uM)

0.8 ul

400 nM

Probe (10 uM)

0.4 ul

200 nM

Template DNA

human genomicDNA:1 to 29 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

95 °C

2 min

Polymerase activation, use 2 minutes for cDNA and 3 minutes for genomic

 

95°C

15 seconds

Denaturation

 

55°C to 60°C

60 seconds

Anneal/Extension do not exceed 60 seconds ; initial experiment use 57 °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

Manufacturer

Instrument

Lo-ROX

Hi ROX

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

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