GC content: As with primer sequences, aim for a GC content of 35–65% and avoid a G at the 5’ end to prevent quenching of the 5’ fluorophore. Primer and probe design considerations Complementarity and secondary structure: Primer and probe designs should be screened for self-dimers, het...
The downside of the SYBR Green assay is that the dye binds to any double-stranded DNA sequence. This means that you could also detect fluorescence emitted from non-specific qPCR products, such as primer dimers. To eliminate this risk, check the reaction specificity by performing a melting curve...
Another reason for incomplete digestion of PCR fragments may be primer dimers. If the restriction site is built into the primer, primer dimers will contain a double-stranded version of the site, usually in vast molar excess over that of the desired target PCR fragment. This problem can be ...
primer annealing temperature, then be lowered by 1-2 °C every second cycle.16These high temperatures during the first cycles avoid PCR primers forming primer-dimers or binding to regions outside the DNA sequence of interest. The downside is that the PCR primers don't all sufficiently bind ...
Processing of genome 5’ termini as a strategy of negative-strand RNA viruses to avoid RIG-I-dependent interferon induction PLoS One, 3 (2008), p. e2032 CrossrefView in ScopusGoogle Scholar [28] H. Kato, et al. Differential roles of MDA5 and RIG-I helicases in the recognition of RNA...
For example, Mason-Pfizer Monkey Virus (MPMV) uses a structured RNA element termed constitutive transport element (CTE) that employs the cellular NXF1/Tap export pathway to avoid nuclear retention of intron-containing RNAs (Bray et al., 1994). This pathway is Crm1-independent, as treatment ...
Another emerging idea about how to prevent AD is rather to avoid the formation of toxic oligomers. Therefore, instead of looking for fibrillization inhibitors, synthetic peptides were designed to trap Aβ1–42 in chimeric amyloid-like fibrils [70]. In addition, Nardo et al. [71] studied the...
Another reason for incomplete digestion of PCR fragments may be primer dimers. If the restriction site is built into the primer, primer dimers will contain a double-stranded version of the site, usually in vast molar excess over that of the desired target PCR fragment. This problem can be ...