Assay system with a single labeled primer containing an iC with a natural reverse primer. During PCR amplification, iGTP-quencher is specifically incorporated opposite the iC, causing a dramatic and rapid decrease in fluorescence as shown in the bottom panel. Multiplexing is achieved by the use of multiple dyes that do not spectrally overlap.
Step 3
Post PCR thermal melt Control for correct amplified sequence Long shelf life
The unique characteristics of EraGen technology were also used to develop MultiCode, a high throughput genotyping system (Figure 4). Overall, the advantages of the Multi-Code multiplexing platform are as follows:
Flexibility No washing, no centrifugation, no filtration High sensitivity and specificity High multiplexing capability Implemented on Luminex instrumentation
The three steps of the Multi-Code system are shown below. Step one is standard multiplexed PCR using a primer with an iC at the 5′ end in the absence of iG-biotin, which results in a “gated” amplicon. Primer extenders and iG-biotin are added for the allelic-specific primer extension step, which results in extended primers with a label and specific capture code. In the third and final step, SAPE and Luminex microparticles are added just prior to injection onto the Luminex instrument. Data analysis is performed on EraGen’s proprietary software.
Summary The EraGen Technology is being implemented in some of the most impressive new platforms to hit the market recently. This new paradigm shift in fundamental biology will allow scientists to greatly simplify genetic analysis.1374639-75-4 medchemexpress With these new reagents and their improved characteristics, EraGen Biosciences is proud to be working with Glen Research to provide today’s researcher with exciting new alternatives in molecular diagnostics and genetic analysis tools. For more information please visit eragen or reference recent publications:
Nucleic Acids Research, 2003, 31(17), 5048-5053, and Clinical Chemistry, 2003, 49(3), 407-414.
Employing EraGen technology in this fashion allows all Multi-Code assays to be performed in a single reaction vessel requiring less plastic and allowing for easier automation.2171061-85-9 Description
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ACTIVATORS, COLUMNS AND PLATES
Benzylthiotetrazole (BTT) first activator described for phosphoramidite chemistry was 1Htetrazole (1), which has served well over the years.PMID:31194385 Nevertheless, more potent activators, 5-methylthio-1H-tetrazole (2) and 5nitrophenyl-1H-tetrazole (3), have popped up but fallen down. More recently, 5ethylthio-1H-tetrazole (ETT) (4) has gained considerable acceptance as a more acidic activator. ETT has the added advantage of being more soluble in acetonitrile than 1Htetrazole (up to 0.75M versus 0. 5M solution in acetonitrile). Less acidic but much more nucleophilic is 4,5-dicyanoimidazole (DCI) (5). DCI is even more soluble in acetonitrile (up to 1.2M solution in acetonitrile). ETT and DCI have proved popular for high throughput synthesizers since they do not tend to crystallize and block the fine outlet nozzles. The renewed interest in RNA synthesis due to the explosion of siRNA technology has led us to evaluate 5-benzylthio-1Htetrazole (BTT) (6), which was described1 several years ago as an ideal activator for RNA synthesis using TOM-protected RNA phosphoramidites2 and recently3 for TBDMSprotected monomers. Our experiments with BTT have shown that its maximum solubility is about 0.33M in acetonitrile The optimal coupling time for TOM-protected RNA phosphoramidites was found to be 90 seconds and for TBD.MedChemExpress (MCE) offers a wide range of high-quality research chemicals and biochemicals (novel life-science reagents, reference compounds and natural compounds) for scientific use. We have professionally experienced and friendly staff to meet your needs. We are a competent and trustworthy partner for your research and scientific projects.Related websites: https://www.medchemexpress.com
