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Custom DNA Probes

KiCqStart™ Primers Frequently Asked Questions

Product No. KSPQ12012

What does validated in silico mean?
What makes KiCqStart™ Primers MIQE compliant?
What is the typical amplicon size?
Do KiCqStart Primers detect all splice variants?
How do I find out the targeted splice variant?
How do I find out the specific splice variant?
Do KiCqStart Primers work in probe-based RT-qPCR?
Can KiCqStart Primers be used to detect genomic DNA?
Where can I find the sequence information?
Do KiCqStart Primers come with any other reaction components, e.g. buffer?
Will the primers for my human gene work with the mouse homologue?
During the design process, which ‘omes’ did off-target searches reference?
During the design process, were amplicons checked for secondary structure?
How do I cite the product in publications?

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 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) and refer to us as Sigma Life Science. Let us know when you publish, and we will send you a gift.