Therefore, additional cDNA cloning strategies are required to confirm the short tag expression3. Conventional Sanger DNA sequencing has been widely used in gene expression profiling and genome sequencing projects. For large-scale transcriptome sequencing, however, the Sanger sequen- cing method is too ...
Sanger sequencing of exons 2-11, and flanking intron regions of the TP53 gene was performed. HapMap population frequencies for TP53 polymorphisms were used as reference values.Results: Five of 90 patients in our study were found to harbor known deleterious mutations in TP53, consistent with LFS....
Elute in 16 µl EB. 2 µl for nanodrop 1 µl for Bioanalyzer ? µl for subcloning and Sanger sequencing (optional) 15. If subcloning: use Invitrogen pCRII-Blunt Topo cloning kit. Try for 3:1 ratio of insert:vector for ligation. Sequence ~ 25 colonies per sample. ...
Sequencing was performed on a Sanger 3500xL Genetic Analyzer (Applied Biosystems™). All samples were measured in triplicates. Primers can be found in Supplemental Table S3. 2.7. RNA Isolation and miRNA qRT-PCR Samples were lysed in TRIzol (Life Technologies, 15596-026, Waltham, MA, USA),...
DNA sequencing, DNA amplification by polymerase chain reaction (PCR), and gene expression by rt-PCR, is of great importance for industrial scaling. However, molecular typing requires the extraction of DNA in good quality and quantity, which is quite difficult to extract. Although several methods ...