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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.

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.

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.

What purification scale is sbeadex Lightning suitable for?

SBXLCK, SBXLCKSTART

The standard protocol assumes a 200 µL volume of lysate for each purification. The purification protocol can be upscaled or downscaled depending on laboratory requirements. We do not provide different scales of kits (e.g. mini, maxi); instead core kit reagents and additional reagents can be adjusted to suit your specific laboratory process.

Are there specific recommendations for performing 1-step RT-qPCR on RNA isolated from clinical samples?

PCRAMP-021

The nucleic acid (either DNA or RNA) will most likely need to be purified from clinical samples due to sample/transport media specific inhibitors and RNAses. For example, for RNA purification, we recommend our sbeadex‚Ñ¢ Pathogen Nucleic Acid Purification Kit for optimal purification.

Should I use electrocompetent or chemically competent cells to make my library?

We recommend using electrocompetent cells for library generation, because the transformation efficiency for electrocompetent cells is higher. Higher transformation efficiency preserves library diversity.

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

AMPSEQKIT1
Currently, we are unable to support human clinical diagnostic applications.

Can I use other dyes besides the Green Fluorescent Dye available in the LavaLAMP™ DNA Component Kit with Dye?

Yes. Syto-13 (Molecular Probes) and EvaGreen (Biotium) have both been tested with the LavaLAMP DNA Master Mix and they perform well.

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.

Do you have any recommendations for purifying DNA from difficult biological samples such as plant leaves and seeds before shearing?

NXGSEQ-003, NXGSEQ-004, 15300-1, NXGSEQ-001, NXGSEQ-002

No. We do not have any specific recommendations for DNA purification. However, the purified DNA should be high quality with A260/280 ratios above 1.8. The DNA should also be free of detectable RNA contamination. Prior to shearing/fragmentation, the purified DNA samples can be analysed on 0.7% agarose gel to check for sample degradation and RNA contamination.

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