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Are there any limitations on the types of fluorescent reporter molecules I can use with RapiDxFire qPCR 5X Master Mix GF?

PCRAMP-012

All fluorescent dyes listed on our Fluorophore and BHQ Selection Chart are available for use with RapiDxFire qPCR 5X Master Mix GF, in which the corresponding BHQ dye has been identified.

Can intercalating dyes be used with this mix?

BHQ-001

BHQ Probe Master Mix has been optimised for use with hydrolysis-probe based chemistry, for both end-point genotyping and qPCR applications. For use with intercalating dyes, we would recommend that you perform your own optimisation and validation to ensure reproducible sensitivity and specificity is achieved using your selected dye.

Are there any limitations on the types of fluorescent reporter molecules I can use with BHQ Probe Master Mix?

BHQ-001

All fluorescent dyes listed on our Fluorophore and BHQ Selection Chart are available for use with BHQ Probe Master Mix, in which the corresponding BHQ dye has been identified.

Can Amp-Seq technology be used in human clinical diagnostics?

AMPSEQ-001
Currently, we are unable to support human clinical diagnostic applications.

Does Amp-Seq provide data analysis software to generate genotype calls?

AMPSEQ-001
Biosearch Technologies provides an analysis pipeline (the BiosearchCaller software), compatible with the Amp-Seq technology, to produce genotypes from the sequencing data. The software can be accessed via a cloud system and our technical team will support the deployment and initial setup.

Why are genotype calls not achieved for specific targets?

AMPSEQ-001
Sometimes, despite a successful primer design for a target, genotyping calls are not achieved for a specific target. This can be caused by several issues including poor primer design and issues with the synthesis of the primers. In addition, if genomic reference data for both parental strains is not provided for target primer design, some primer pairs might not work for all F1 samples. Missing genotypes are generally attributed to low depth. Certain targets cannot be amplified because of sample quality issues or unknown parental bias in the genomes. Our standard design process addresses the parental bias by avoiding primers on known variability in the genome. Known variation information can be provided to us in the target flanking sequences through the Data Submission form.

Why are genotype calls not achieved for specific targets?

AMPSEQKIT1

Sometimes, despite a successful primer design for a target, genotyping calls are not achieved for a specific target. This can be caused by several issues including poor primer design and issues with the synthesis of the primers. In addition, if genomic reference data for both parental strains is not provided for target primer design, some primer pairs might not work for all F1 samples.

Missing genotypes are generally attributed to low depth. Certain targets cannot be amplified because of sample quality issues or unknown parental bias in the genomes. Our standard design process addresses the parental bias by avoiding primers on known variability in the genome. Known variation information can be provided to us in the target flanking sequences through the Data Submission form.

There are transformation protocols for chemically competent cells that do not include a heat shock. Do I need to heat shock my competent cells?

We have not tested alternative transformation protocols extensively. You can avoid the heat shock step, but it is highly likely that your transformation efficiencies will decrease.

What if I don’t have a fluorimeter? How else can I monitor the reaction?

Amplified products can also be monitored by viewing the reaction on an agarose gel or by measuring the turbidity of the reaction at OD600.

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