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ADVANCEDtm qPCR HRM mix

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ADVANCED™qPCR HRM Mix
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介绍详情

Ordering Information:

CAT.NO.

Quantity

Specification

801-030-AR

100 REACTIONS

HRM MIX

801-030-ER

500 REACTIONS

HRM MIX

801-030-TR

2000 REACTIONS

HRM MIX

For instrument compatibility refer to APPENDIX II

PRODUCT DESCRIPTION:

High Resolution Melt (HRM) analysis is a powerful technique for the analysis of mutations, polymorphisms and epigenetic differences in double-stranded DNA samples.

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OUR MIX OFFERS:

 Accurate SNP genotyping

Gene scanning

CpG methylation analysis

3rd generation, saturating SuperGreen

Efficient specific amplification from GC rich and AT rich sequences

ADVANCED HRM Mix uses the novel SUPERGREEN intercalating dye does not inhibit PCR and is a saturating, dye essential for accurate SNP genotyping. SUPERGREEN can be used at high concentrations and does not have a different affinity for AT or GC rich sequences.

Product Stability And Storage:

On arrival store at -25°C to -15℃.

Avoid prolonged exposure to light.

If stored correctly the mix 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 HRM mix .

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

Add

Volume

Final concentration:

WISENT ADVANCED HRM mix

10 ul

1X

Forward primer (10 uM)

0.8ul

400nM

Reverse primer (10 uM)

0.8ul

400nM

Template DNA

cDNA : < 100ng
genomic DNA: <1ug

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

cDNA:2min
genomic DNA:3min

Polymerase activation

40

95°C

5 seconds

Denaturation

60°C to 65°C

20-30 seconds

Anneal/Extension, do not exceed 30 seconds, do not use temperatures below 60°C

HRM analysis

Refer to instrument instructions

optional

APPENDIX II 

Manufacturer

Instrument

Applied Biosystems

7900, 7900HT, 7900HT FAST, StepOneTM, StepOne TM Plus, 7500,
7500 FAST, Viia7tm

Bio-Rad®

CFX96TM, CFX384TM

Eppendorf

Mastercycler® ep realplex, Mastercycler® realplex 2S

Illumina®

EcoTM

Qiagen/Corbett

6000, Q

Roche Applied Science

Lightcycler®480, Lightcycler®Nano

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