Select your products and services :
Select your country :
Americas
Europe
Asia and South Pacific
Middle East and Africa

Update your choices | BU : Life Science | Country : USA

Key factors to select the right qPCR kit

key factors to select your qPCR kit

Several parameters have to be taking into to account to select the right kit for your qPCR assays. Here, we explain whys and wherefores of crucial and optional parameters.

qPCR Selector

An online tool has been created to help you in your kit selection, the qPCR selector!

Crucial parameters in your qPCR kit selection

Detection method: SYBR® vs Probes

In order to detect and measure the amount of target in the sample, a measurable signal has to be generated, which is proportional to the amount of amplified product. All current detection systems use fluorescent technologies.

SYBR®

SYBRR Green I is the most commonly used dye for non-specific detection. It is a double-stranded DNA intercalating dye, that fluoresces once bound to the DNA. The amount of dye incorporated is proportional to the amount of generated target.

Probes

Alternatively the signal can be generated using an additional oligonucleotide, harbouring a fluorescent dye and a quencher. This also ensures a higher specificity to the assay.

 
Discover our range of qPCR Probes   >
 

RT-qPCR: one-step vs two-step reactions

Takyon™ kits are intended for the amplification of DNA by qPCR but kits are also available for RT-qPCR assays. If your starting material is RNA, you have to perform the Reverse Transcription (RT) to get cDNA material before starting your qPCR assay. This can be done in a One-Step or a Two-Step approach. The One-Step Takyon™ MasterMix contains the new EuroscriptII RT which delivers highly robust performance on classical mRNAs, small RNA and viral RNAs. The new additive improves significantly the amplification performance on some difficult virus types as well as on FFPE samples. A Two-Step reaction consists in separate RT and qPCR (with MasterMix or Core Kit) reactions

One-Step RT converter

Any Takyon™ dTTP Master Mix for qPCR assays can easily be converted into a one-step RT-qPCR kit by using the Takyon™ One-Step kit converter.

Normalisation: ROX, low-ROX, no-ROX

Some thermocyclers require internal ROX dye for normalisation. ROX is an inert additive acting as a passive reference dye that gives a constant signal for well-to-well normalisation in a qPCR assay. Working with the ROX dye already incorporated in the MasterMix guarantees a higher level of reproducibility and homogeneity to your assays when compared to manual addition of ROX.

Optional parameters in your qPCR kit selection to fine tune your assay

Format: Mastermix vs core kits

MasterMix

A MasterMix is a ready-to-use reagent that contains all the qPCR components in a unique vial, ensuring maximal reproducibility of results with a minimal contamination risk. Additionally to the 2X MasterMixes format, the 5X highly concentrated kits allow template volume optimisation in the context of small volume reaction or of poorly concentrated templates. Our MasterMixes are available for SYBRR and Probe Assays (see Detection method).

Core kit

Beside the MasterMixes, our Core Kits are cost effective solutions that offer maximum flexibility as you receive the different components of the MasterMix in separate tubes, allowing you to maximise assay performance through the optimisation of component concentrations.

Carry over: dTTPs vs dUTPs

Some kits contain a blend of dNTPs and dUTP with UNG, other ones contain only dNTPs. Using only dNTPs increases the sensitivity, the reason being that the Taq® DNA polymerase incorporates more easily dTTPs than dUTP. However, using a mix containing dUTP and UNG brings security to the assay. Indeed any trace of DNA molecule from previous reaction containing dUTP will be removed by the action of the Uracyl-N-Glycosylase (UNG). The Uracil-N-Glycosylase (UNG) is an enzyme that hydrolyses all singlestranded and double-stranded DNA containing dUTPs. Consequently, if all PCR amplifications are performed in the presence of a dNTPs/dUTP blend, by carrying a UNG step before every run it is possible to get rid of any previous undesired PCR product containing dUTP.

Visual aid

The inert blue dye allows you to visualize dispensing errors at a glance and avoid aberrant results before they occur. This is particularly true when using white plates, known for their superior performances.

Assay Atomation

The low viscosity HTP Takyon™ kits are perfectly suited for assay set-up automation.



qPCR Range

Discover our Takyon™ qPCR range: You can evaluate a sample of any MasterMix for free and benefit from a 50% discount on your first kit order.



Support Area


We use cookies to ensure the best experience on our website. By continuing to use this website, you consent to our cookie policy.