Skip to content Skip to navigation menu
cancel
  1. Home
  2. BHQ dye

You searched for "BHQ dye"

  • 1
  • 2(current)
  • 3

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.

What are the storage conditions for BHQ Probe Master Mix?

BHQ-001

BHQ Probe Master Mix should be stored at -20 °C. Repeated freeze-thaws should be avoided, so if BHQ Probe Master Mix is to be aliquoted, nuclease-free, light-protected tubes/vials should be used.

Once thawed, BHQ Probe Master Mix can be stored at +4 °C for 1 week.

Can BHQ Probe Master Mix be used in multiplex reactions?

BHQ-001

Yes, it has been optimised for use in multiplex qPCR applications and customers can chose from a wide range of fluorophores from Biosearch Technologies, depending on their qPCR instrument.

When performing multiplexing reactions, we recommend optimising each assay as a singleplex reaction, before combining a multiplex, to evaluate the optimal parameters of each assay (e.g. annealing temperature, cycling conditions etc.)

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 BHQ Probe Master Mix be used for both end-point genotyping and qPCR applications?

BHQ-001

BHQ Probe Master Mix is a highly versatile PCR Master Mix, which has been validated for use with both endpoint genotyping and qPCR applications. Please refer to the BHQ Probe Master Mix User Guide for detailed instructions and protocols.

What concentration of primers and probes should I used in my reactions?

BHQ-001

Please refer to the BHQ Probe Master Mix User Guide for detailed information and recommended protocols.

We recommend a final concentration of 900 nM of primer and 200 nM probe in each reaction for qPCR applications. Please note, that for multiplex reactions, further optimisation may be required.

If using BHQ Probe Master Mix for endpoint genotyping applications, we recommend final concentrations of 400 nM primer and 200 nM probe as starting concentrations during initial optimisation.

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.

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.

  • 1
  • 2(current)
  • 3