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 assays have undergone wet-lab validation experiments and all performed well.
What makes KiCqStart® Probe Assays MIQE compliant?
Among other things, the exon locations, sequences, and quantities of the oligonucleotides are provided either before or after purchase.
Do KiCqStart® Probe Assays detect all splice variants?
No, the assays have been designed to detect the most prevalent one for each eukaryotic gene based on a literature review. However, it is possible an assay may co-amplify other transcripts.
How do I find out the targeted splice variant?
Search GenBank with the assay RefSeq.
How do I find out the specific splice variant?
Search GenBank with the assay RefSeq.
Do KiCqStart® Probe Assays work in SYBR® Green I RT-qPCR?
Possibly, but the forward and reverse primers have been specifically designed for probe-based detection. For SYBR Green I assays, please try KiCqStart® Primers.
Can KiCqStart® Probe Assays be used to detect genomic DNA?
No, the assays have been specifically designed to detect mRNA transcripts and their corresponding cDNA. However, at least some homology is usually unavoidable, so it may occur with select assays.
Where can I find the sequence information?
Sequence information will be provided on the Standard CoA / Technical Datasheets that accompany each assay shipment.
Do KiCqStart® Probe Assays come with any other reaction components, e.g. buffer?
No, only one forward primer, one reverse primer, and a probe, dry, in individual tubes is included.
Will KiCqStart® Probe Assays for my human gene work with the mouse homologue?
Possibly, but the assays 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 the amplicons and / or the primers and probe.
Are other reporter dyes available?
Yes, KiCqStart® Probe Assays can be ordered on a custom basis with other reporter dyes. Depending on the dye, it might be necessary to change the quencher from Onyx Quencher™ A to Onyx Quencher™ B, C, or D. Please contact your local sales professional for assistance.
Are other quenchers available?
No, KiCqStart® Probe Assays come exclusively with Onyx Quencher™.
Are internal modifications available?
No, KiCqStart® Probe Assays contain A, C, G, and T bases only.
Is it possible to order a larger amount of material?
Yes, we can manufacture the assays in larger quantities on a custom basis. Please contact your local sales professional for assistance.
Is it possible to order the assays in a different format?
Yes, we can format the assays on a custom basis, e.g. mixed primers and probe in plates. Please contact your local sales professional for assistance.
How do I cite the product in publications?
The best approach would be to mention the species, gene, gene ID, and assay (1, 2, or 3) and refer to use as MilliporeSigma. Let us know when you publish, and we will send you a gift.
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