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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.

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.

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 Endura High-Yield Electrocompetent Cells classified as genetically modified organisms (GMO)?

COMCEL-017
Depending on the definition of GMO, yes, they are GMO as they were constructed by methods outside of E. coli conjugations or viral transductions. However, they do not contain DNA from organisms other than E. coli.

Have you produced proteins at scale?

COMCEL-008

Our customers have reported growth of ClearColi strains in a 5 litre fermenter and shown optical density levels in excess of 150, and that some defoaming agents (particularly silicone-based defoamers) are beneficial while other defoaming agents such as polypropylene glycol may inhibit cell growth.

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.

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.

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.

How do I calculate transformation efficiency (TE)?

TE = (# colonies) / (µg DNA transformed) * (final transformation volume) / (volume plated) * dilution factor = cfu / µg

For example, if you transformed 1 ng of plasmid DNA into cells, diluted the transformation to a final volume of 1000 µL with Recovery Medium, then plated 25 µL of a 1:100 dilution of that transformation, and the next day you counted 150 colonies on your plate, your calculation would look like this:

# of colonies: 150
µg DNA transformed: 0.001
Final tansformation volume = 1000 µL
Volume plated = 25 µL
Dilution factor = 1 µL into 100 µL, 100-fold (factor of 100)

TE = (150 / 0.001) * (1000 / 25) * 100 = 6 x 108 cfu / µg

How do you determine the brightness of a dye?

The absolute intensity of a dye is a product of the extinction coefficient and the quantum yield. We have not measured the quantum yield for our dyes as this value is highly dependent upon the local environment, including the buffer system used for the measurement. However, we do provide the extinction coefficients for dye modifications at their lambda max wavelength, and these values are available under the Technical Specs tabs of our Oligo Modifications webpages. While quantum yield and extinction coefficients both contribute to dye detectability, the principal determinant for Stellaris® RNA FISH assays is actually the instrument optics, including the excitation source, available filters, and quantum efficiency of the camera.

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