TATAA two-tailed miRNA assays

TATAA two-tailed RT-qPCR microRNA assays
Our new TATAA miRNA two-Tailed RT-qPCR assays show exceeding performance compared to other techniques as recently published in NAR (see Product Documents).

The challenge detecting small microRNAs is that two conventional PCR primers do not fit the target as their combined length is almost twice the length of the microRNA. Older techniques have solved this by extending the microRNA using eg., a hairpin primer, adding a poly A-tail, or adding a fragment by ligation. This, however, compromises the assay sensitivity and specificity, as only one of the PCR primers sense the actual microRNA sequence; the other senses the added extension. Further, these methods fail to detect microRNAs modified in the 3’-end as it interferes with the extension process.

TATAA miRNA two-Tailed RT-qPCR assays offers a superior solution. Instead of using a single binding probe, Two-tailed PCR uses two hemiprobes, which bind to different stretches of the microRNA, that are connected by a folded tether. While each hemiprobe is too short to bind the microRNA, when both hemiprobes are complementary they bind cooperatively.

Advantages
Binding is exceeding specific, as a mismatch is much more profound in a short hemiprobe
The cDNA formed can then be PCR amplified using two sequence specific primers
SYBR used for detection
High degree two-tube multiplexing of the RT followed by singleplex qPCR

Contents
100 ul (2 uM) RT primer
100 ul (10 uM) qPCR primer

Assays currently available

miR-23a-3p (AUCACAUUGCCAGGGAUUUCC)
miR-451a-5p (AAACCGUUACCAUUACUGAGUU)
cel-miR-39-3p (UCACCGGGUGUAAAUCAGCUUG)
miR-1909-3p (CGCAGGGGCCGGGUGCUCACCG)
miR-5739 (GCGGAGAGAGAAUGGGGAGC)
miR-5001-5p (AGGGCUGGACUCAGCGGCGGAGCU)
miR-3185 (AGAAGAAGGCGGUCGGUCUGCGG)
miR-6795-5p (UGGGGGGACAGGAUGAGAGGCUGU)
miR-122-5p (UGGAGUGUGACAAUGGUGUUUG)
miR-16-5p (UAGCAGCACGUAAAUAUUGGCG)
miR-199a-3p (ACAGUAGUCUGCACAUUGGUUA)
miR-21-5p (UAGCUUAUCAGACUGAUGUUGA)

Customized Assays
To place a customized assay order, Contact us.

Two-tailed RT-qPCR assay is designed for microRNA. The assay is validated on RT and qPCR level on synthetic microRNA.

Below, links to independent papers developing and using Two-Tailed PCR can be found

https://www.frontiersin.org/articles/10.3389/fpls.2019.01234/full
https://www.mdpi.com/1422-0067/20/20/5131/htm#app1-ijms-20-05131

Click here for more papers citing Two-tailed PCR

Two-Tailed PCR presentations:

Dr Kubista presents Two-Tailed PCR at GQ2019: Click here>

Biovendor presentation on Two-Tailed PCR: Click Here>