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Can BHQ Probe Master Mix be used with crudely-extracted DNA?

BHQ-001

Yes, BHQ Probe Master Mix has been optimised to with crudely-extracted DNA (Biosearch Technologies QuickExtract, HotShot) on a variety of agrigenomics samples (e.g. seeds, leaves), as well as on purified DNA. If working with agrigenomics samples, it is recommended to use dry material (e.g. desiccation of leaves using the Biosearch Technologies Plant Sampling Kit). This will allow for the most optimum recovery of DNA. Please contact our Technical Support team for further information.

What reaction volumes can be used with BHQ Probe Master Mix?

BHQ-001

BHQ Probe Master Mix has been validated for use in endpoint genotyping and qPCR applications in 1.6 µL – 25 µL reaction volumes.

What are the optimum thermal cycling conditions when working with BHQ Probe Master Mix?

BHQ-001

The thermal cycling conditions will depend various factors. For example, the annealing temperature will depend on the sequence of the target-specific primers and probes used in the reaction, and the number of cycles will be dependent on the efficiency of the PCR reaction. Please refer to the BHQ Probe Master Mix User Guide for further information on recommended protocols and adhering to the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) Guidelines.

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

PCRAMP-009, PCRAMP-010, PCRAMP-007, PCRAMP-006

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

What is the difference between RapiDxFire™ qPCR 5X Master Mix GF and BHQ™ Probe Master Mix?

PCRAMP-012

RapiDxFire qPCR 5X Master Mix GF is a 5X qPCR master mix produced and purified in a quality standard ISO 13485-certified manufacturing facility, allowing for its use in molecular diagnostic and clinical applications.

BHQ Probe Master Mix is a 2X PCR master mix, which has been validated for both endpoint genotyping and qPCR applications. It has been designed specifically to be inhibitor-tolerant, especially when working with crude-extracted agrigenomic samples (e.g. leaves, seeds).

Does the RapiDxFire qPCR 5X Master Mix GF contain a passive reference dye?

PCRAMP-012

No, the RapiDxFire qPCR 5X Master Mix GF does not contain a passive reference dye. However, customers can supplement their reaction mix with a passive reference dye of their choice.

Is BHQ Probe Master Mix stable at room temperature?

BHQ-001

BHQ Probe Master Mix is an incredibly stable PCR/qPCR Master Mix, which shows little-to-no reduction in performance when left at room temperature, even when combined with all other PCR-reaction components. We have demonstrated Cq > 1.0 when assembled qPCR reactions were left at room temperature for 16 hours, in both singleplex and multiplex applications.

To maintain high performance, it is recommended to store BHQ Probe Master Mix at -20 °C, and aliquot it into nuclease-free, light-protected tubes to minimise the number of freeze-thaw cycles. BHQ Probe Master Mix is also stable at +4 °C for 1 week.

What is the difference between lipid IVA and LPS?

BHQ-001

Lipid IVA is a modified LPS molecule that does not induce an endotoxic response in human cells. The six acyl chains of LPS (Figure 1) that trigger the endotoxic response are recognised by Toll-like receptor 4 (TLR4) in complex with myeloid differentiation factor 2 (MD-2), causing NF-?B activation and production of proinflammatory cytokines.  Lipid IVA lacks the two secondary acyl chains and does not induce formation of the activated hTLR4/MD-2 complex, thus evading the endotoxic response.  In addition, the oligosaccharide chain is deleted, making it easier to remove the resulting homogenous lipid IVA from any downstream product.

ClearColi lipd IVA and LPS from unmodified E. coli

Figure 1:  Comparison of ClearColi lipid IVA and LPS from unmodified E. coli.  In ClearColi, two of the six acyl chains have been removed to disable the endotoxin signal, and the oligosaccharide chain has been deleted.

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

PCRAMP-009, PCRAMP-010, PCRAMP-007, PCRAMP-006

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

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