• I'm using a website: Northwestern university/ biotools that shows if your primers have any complementarity and unfortunately two of my forward primers happen to be complementary at a particular stretch. (protocol-online.org)
  • 2010). In the RF method, PCR primers are replaced with long dsDNA that has 5' ends containing homologous overlaps with the desired vector insertion site (Figure 2). (idtdna.com)
  • Unlike kits that rely on linear amplification, primers designed for the Q5 Site-Directed Mutagenesis Kit should not overlap to ensure that the benefits of exponential amplification are realized. (neb.com)
  • Since 2013, the development of the CRISPR/Cas9 technology, based on a prokaryotic viral defense system, has also allowed for the editing of the genome, and mutagenesis may be performed in vivo with relative ease. (wikipedia.org)
  • We further demonstrate the ease with which mutagenesis can be achieved by deleting 13.5 kb from the Toledo genome using a PCR-based technique. (biomedsearch.com)
  • To evaluate whether Cpf1 can be applied to plant genome editing, we selected Cpf1 from Francisella novicida (FnCpf1), which recognizes a shorter PAM (TTN) within known Cpf1 proteins, and applied it to targeted mutagenesis in tobacco and rice. (nature.com)
  • In order to develop a more "human-like" uricase for therapeutic use, exon replacement/restoration and site-directed mutagenesis were performed to obtain porcine-human uricase with higher homology to deduced human uricase (dHU) and increased uricolytic activity. (mdpi.com)
  • Docking of RP 73401 into the active site of a 2B6 homology model suggested a direct interaction with residue L363 but not with F107. (aspetjournals.org)
  • Previously in our laboratory, eight highly active OmpT variants were engineered with novel catalytic sites. (utexas.edu)
  • Hydrophobic interactions and shape complementarity enhance peptide affinity beyond the catalytic site and support a limited selectivity model for antigenic peptide selection that may underlie the generation of complex immunopeptidomes. (jimmunol.org)
  • The Big Blue transgenic rodent mutagenesis assay is an in vivo system for assessing the genetic toxicity of test compounds. (selectscience.net)
  • In addition, two Sp1 binding sites in HS4 were also found to positively influence bcl-2 activity, and Sp1 was observed to interact with the human HS4 enhancer in vivo . (aacrjournals.org)
  • No activity was detected after the histidine residue at the phosphorylation site of the alpha-subunit was replaced by aspartate (His-246 alpha----Asp), but this alteration seemed to have a deleterious effect on the accumulation of the enzyme in cell-free supernatant extracts. (portlandpress.com)
  • Comparison of available subunit I sequences reveals that the only structural elements conserved within the oxygen reductase families that could perform these functions are active-site components, namely the covalently linked histidine-tyrosine, the Cu B and its ligands, and the active-site heme and its ligands. (pnas.org)
  • The K-channel leads from a glutamate residue at the cytoplasmic interface of subunits I and II to the cross-linked histidine-tyrosine cofactor in the active site (see ref. 6 for review). (pnas.org)
  • Substitutions within and deletion of the entire active-site-occluding loop demonstrated a prominent role for His-44 and this loop in the structure and activity of CT. (asm.org)