PCR-based strategy was used to construct the inducible NCD biosynthesis module (Supplementary Table 7). Primers pA-PolB-294 and pA-DedA-228 were used to amplify the ara operon from E. coli genome, and the fragment was inserted into pUC18 to generate pUC-PolB-DedA. The original genes araB...
When these PCR products are mixed, denatured and allowed to anneal, the complementary ends of the DNA hybridize, serving as primers for the extension of the complementary strand. The result is that the two genes are spliced together. This fusion gene can then be amplified by a second PCR. ...
and pose significant threats to human health and structural integrity problems in built environments. While most fungal diversity remains unknown, the development of PCR primers for the internal transcribed spacer (ITS) combined with next-generation sequencing has substantially improved our ability to...
(1994) Solid phase sequencing of PCR products. In: McPherson MJ, ed. PCR II - A Practical Approach. Oxford: IRL Press, Oxford University Press. Nelson WM, Wojnar WA (1991) The use of photobiotinylated PCR primers for magnetic bead-based solid phase sequencing. Human...
Although the QuikChange was originally developed for site-directed mutagenesis using complementary mutagenic oligonucleotide primers in whole plasmid PCR, we found that the protocol also worked for megaprimers consisting of hundreds of nucleotides. Based on this discovery, we used insert fragments, ...
During the annealing step of a qPCR assay, the probe and PCR primers attach to their complementary sequences in the target region. Then, during primer extension or elongation, the exonuclease activity of DNA polymerase degrades the probe, releasing the 5’ fluorophore from the quencher. Without...
Here, we developed a novel strategy to create tumor antigens by construction of random tumor transcriptome expression library (RTTEL). The complementary DNA (cDNA) from S180 sarcoma was used as template for arbitrarily amplifying gene fragments with random primers by PCR, then ligated to the C-...
Two pairs of primers, specific for genomes of BY4742/BY4742a and SY14/SY14a, were used. Data shown are representative images of two independent experiments. Extended Data Table 1 Details of the creation of a single chromosome yeast Full size table Extended Data Table 2 Information regarding ...
For PCR reactions that did not yield products, quality check was carried out by a new round of PCR with kojR primers whose PCR product was 1.0 kb. PCR products were purified by a DNA Clean & Concentrator-5 kit (Zymo Research) and sequenced at the Iowa State University DNA Facility (...
The PCR primers shown in Table I were designed to provide specific-restriction sites at the ends so that the amplified product could be cloned directly into the vector plasmid into plasmid pRSET-B. The plasmid carrying an integrated CA is referred to as R1. ...