Using a plastic sheet (overhead transparency) to separate the image and the gel, lay the agarose gel onto the image and excise the smeared library from the region above the primer dimers, which can be identified in the control library. 4. Transfer the excised gel fragment to a clean 1.5...
45 pairs of gene-specific qPCR primers were experimentally validated by SYBR Green method using first-strand cDNA and ddH2O as template. There were no non-specific amplification and primer-dimers produced by melting curve analysis and agarose gel electrophoresis detection. We used a dilution series...
(requiring lengthy primer of high melting temperature and high GC content, and these tend to yield primer dimers and/or strong secondary structures), to the PCR reaction itself (very prolonged reactions, very low reaction yields that usually cannot be distinguished on gel, potential occurrence of ...
We also explored the template specificity of piPolB TLS capacity with substrates containing thymine-glycol (Tg)-oxidized base and cyclobutane thymine dimers (T:T). Our results indicated that piPolB was able to bypass Tg in the presence of magnesium ions (Figure 3D, lanes 5–7). However, ...
These circumstances can lead to self-dimers or primer-dimers instead of annealing to the desired DNA sequences. If you are using the primers for cloning, we recommend cartridge purification as a minimum level of purif...
The final case occurs when the primers interact with each other at the 3' end of either primer to produce primer-dimers (see figure 1C). Figure 1 Primer secondary structure formation. Shown in figure 1A) is the secondary structure stem loop formation for the primer ACATACTGTGAGAAACACAGTA...
* Non-specific amplification and absence of primer-dimers are ensured when All-in-One validated PCR primers and PCR mix are used together. † Primers validated automatically for human, mouse and rat. Inquire for validation of other species. ...
For the failed primer set the forward and the reverse primers were able to form primer dimers and redesign of the primers solved this problem. By taking primer dimer formation into account it may be possible to reach even higher design success rates for DNA primers. In contrast, the ...
V. et al. DNA origami based assembly of gold nanoparticle dimers for surface-enhanced Raman scattering. Nat. Commun. 5, 3448 (2014). ADS Google Scholar Acuna, G. P. et al. Fluorescence enhancement at docking sites of DNA-directed self-assembled nanoantennas. Science 338, 506–510 (2012)...
on standard agarose gel. Our added amplification step (by use of the same short primers initially employed) remedies this limitation and allows to resolve whether the desired Gibson-assembled product has been obtained on agarose gel or by sequencing of amplicons. It also provides large amounts ...