MeltDoctor™ HRM Positive Control Kit

Applied Biosystem® MeltDoctor™ HRM Positive Control Kit provides nucleic acid controls to demonstrate and troubleshoot High Resolution Melt (HRM) analysis.Read more

Catalog Number	Quantity
4410126	1 kit

Catalog number 4410126

Price (CNY)

5,347.00

Add to cart

Quantity:

1 kit

Price (CNY)

5,347.00

Add to cart

Applied Biosystem® MeltDoctor™ HRM Positive Control Kit provides nucleic acid controls to demonstrate and troubleshoot High Resolution Melt (HRM) analysis. The kit consists of forward and reverse primers and three DNA templates representing alleles AA, AB, and BB.

For Research Use Only. Not for use in diagnostic procedures.

Specifications

Control TemplateDNA template representing alleles AA, AB, and BB

FormatTube(s)

Gene SymbolACTB

Kit TypeMeltDoctor HRM Positive Control Kit

No. of Reactions450 Reactions

Product LineMeltDoctor™

Product TypeHRM Positive Control Kit

Quantity1 kit

Shipping ConditionWet Ice

Sufficient For450 Reactions

Targetβ-actin

Volume150 μL, 500 μL

Concentration20X

For Use With (Application)Real Time PCR (qPCR)

Label or DyeHigh Resolution Melting (HRM)

Unit SizeEach

Contents & Storage

The kit consists of: MeltDoctor™ HRM Allele A DNA (20x), 150 μL, MeltDoctor™ HRM Allele G DNA (20x), 150 μL, MeltDoctor™ HRM Allele A⁄G DNA (20x), 150 μL, MeltDoctor™ HRM Primer Mix (20x), 500 μL
Store at -15 °C to -25 °C

Guaranteed minimum shelf life is 60 days (exact expiry date printed on product and CofA).

Frequently asked questions (FAQs)

The HRM software will not accept my calibration file. What is wrong?

Check that there are no outliers on the plate. You cannot omit any wells on the HRM calibration plate.

I have a 7900HT Fast Real-Time PCR System, and cannot open the data file in HRM software. What could be causing this?

There are a few possibilities. First, make sure the HRM Software version is v2.0.1, and the 7900HT Fast Real-Time PCR System software version is v2.3 or above. Second, check that the run method used was as recommended in the HRM protocol; make sure the ramp rate for the dissociation stage is 1%. Then try to open the calibration file from the HRM Software; if it does not open, the calibration file is defective. The defects could be due to a bad calibration plate or instrument uniformity issue.

My 7500 Fast Real-Time PCR System has been upgraded to v2.0.6, so why can't I analyze my files with HRM Software v2.0.1?

For the 7500 Fast Real-Time PCR System, if the software version is below v2.0.4, the compatible HRM Software is v2.0.1. If the 7500 Fast Real-Time PCR System has been upgraded to software version 2.0.4 or above, then you will need to get HRM Software version 3.0.1.

How can I get positive controls for the different genotypes for my HRM experiment?

You can order a synthetic positive control plasmid using our GeneArt Gene Synthesis services. The final product is sequence verified and provided as 5 µg of lyophilized plasmid. Alternatively, you may be able to find a positive control sample from Coriell.

How do I perform the HRM calibration on my real-time PCR instrument?

The HRM calibration is a 3-step procedure. In some instrument software, the calibration process has been streamlined to a single-step procedure (applies to the ViiA7 Real-Time PCR System and QuantStudio Real-Time PCR Systems).

For the HRM calibration, the first step is a PCR reaction to generate the HRM dye plate. The reaction plate contains a DNA template, a primer pair, and the PCR mix with HRM dye. The second step is dye calibration using the PCR plate from step 1. The last step is a melt curve run, again using the same plate. For details, please refer to the HRM guides for each instrument:

- 7500 Fast Real-Time PCR System (with software version 1.4 or 2.0.4 and below) (https://tools.thermofisher.com/content/sfs/manuals/cms_050347.pdf)
- 7500 Fast Real-Time PCR System (with software version 2.0.5 and above) (https://tools.thermofisher.com/content/sfs/manuals/4457847.pdf)
- 7900HT Real-Time PCR System (https://tools.thermofisher.com/content/sfs/manuals/cms_050347.pdf)
- StepOne and StepOnePlus Real-Time PCR Systems (https://tools.thermofisher.com/content/sfs/manuals/4457847.pdf)
- ViiA 7 Real-Time PCR System (https://tools.thermofisher.com/content/sfs/manuals/4443531.pdf)
- QuantStudio 6 and 7 Real-Time PCR Systems (https://tools.thermofisher.com/content/sfs/manuals/4489823.pdf)
- QuantStudio 6 and 7 Pro Real-Time PCR Systems (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0018981_HighResolutionMelt_DA_SW_UG.pdf)
- QuantStudio 3 and 5 Real Time PCR Systems (https://tools.thermofisher.com/content/sfs/manuals/MAN0014394_HighResMeltExperiment_GSG.pdf)
- QuantStudio 12K Flex Real-Time PCR System (https://tools.thermofisher.com/content/sfs/manuals/4470692A.pdf)

Find additional tips, troubleshooting help, and resources within our High Resolution Melt Support Center.

View all MeltDoctor™ HRM Positive Control Kit FAQs