We get multiple bands at very high resolution, and deep bit depth, so we put together this interesting data set of all the interesting things that are gong on right now that we'd like to monitor.
We then attempted to sequence additional SCPCH that yielded multiple bands at the first PCR stage by using PCR touch-downs or gel extraction to isolate the appropriate size band.
However, the PAUs showed multiple bands at 55°C and annealing temperature was increased to 61°C.
3. We did not observe multiple bands at any nucleotide position on sequencing gels as expected if multiple loci were amplified during PCR.
For any sample that showed multiple bands at any of the MIRU loci, the PCR was repeated at least twice in order to confirm the results.
Gelatin zymography showed some induction of MMP-9, as well as multiple bands at around the Mr of the collagenases that are induced by IL-1α/OSM and repressed by the additional presence of HDAC inhibitors in this system.
DOI: http://dx.doi.org/10.7554/eLife.10072.007 When assessing intestinal GSK3βY216 phosphorylation by IB, we detected multiple bands at high molecular weight that might represent phosphorylated GSK3αY279 and/or phosphorylated GSK3βY216 with other types of modifications, possibly ubiquitination.
For any sample that showed multiple bands at any of the MIRU-VNTR loci, the PCR was repeated twice in order to confirm the results as well as to rule out the possibility of contamination.
Notably, multiple bands at different molecular weights were observed by western blotting for FAN1 in M phase, but not in G1 phase, indicating that FAN1 may be phosphorylated in M phase, which is common for mitotic substrates of APC/CCdh1 [27].
Numerous proteins appeared in multiple bands, at different molecular weights (e.g., DMBT1, carbonic anhydrase 1, neutrophilic elastase), suggesting a possible degradation and/or various post-translational modifications in the colon lumen.
