KiCqStart^® SYBR^® Green Primers FAQs

Compatible with any thermal cycler, KiCqStart^® Primers (Product No. KSPQ12012) are ready-to-order, pre-designed primer pairs for quantifying gene expression by SYBR Green I RT-qPCR (Reverse Transcription Quantitative Real-Time PCR). To comply with MIQE, the primers have been developed using sophisticated bioinformatics tools and validated in silico to avoid off-target amplification. KiCqStart® Primers are available as up to three ranked sets of forward and reverse primer pairs for all genes from common model organisms.

What does validated in silico mean?
The performance of the sequences against the targets was verified with computational PCR. Given that the chemistry of PCR is programmed into the software, this ‘computer test tube’ is an excellent surrogate for the expected outcome of any in vitro reaction. In addition, a select number of primer pairs have undergone wet-lab validation experiments and all performed well.

What makes KiCqStart^® Primers MIQE compliant?
Among other things, the exon locations, sequences, and quantities of the oligonucleotides are provided either before or after purchase.

What is the typical amplicon size?
Most amplicons are between 75 and 200 base pairs.

Do KiCqStart^® Primers detect all splice variants?
No, the primer pairs have been designed to detect the most prevalent one for each eukaryotic gene based on a literature review. However, it is possible that the primer pairs may co-amplify other transcripts.

How do I find out the targeted splice variant?
Search GenBank with the primer pair RefSeq.

How do I find out the specific splice variant?
Search GenBank with the primer pair RefSeq.

Do KiCqStart^® Primers work in probe-based RT-qPCR?
No, the primer pairs have been specifically designed for SYBR^® Green I detection chemistry.

Can KiCqStart^® Primers be used to detect genomic DNA?
No, the primer pairs have been specifically designed to detect mRNA transcripts and their corresponding cDNA. However, primer pairs with a ranking of 4 may also co-amplify contaminating genomic DNA.

Where can I find the sequence information?
Sequence information will be provided on the Technical Data Sheets that accompany each primer pair shipment.

Do KiCqStart^® Primers come with any other reaction components, e.g. buffer?
No, only one forward and one reverse primer, dry, in individual tubes is included.

Will the KiCqStart^® Primers for my human gene work with the mouse homologue?
Possibly, but the primer pairs were designed to be species specific.

During the design process, which ‘omes’ did off-target searches reference?
Both transcriptomes and genomes.

During the design process, were amplicons checked for secondary structure?
Depending on the species, it was either the amplicons and / or the primers.

How do I cite the product in publications?
The best approach would be to mention the species, gene, gene ID, primer pair (1, 2, or 3). Let us know when you publish, and we will send you a gift.