Words similar to bamh

Example sentences for: bamh

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Oligonucleotide primers for the PCR reaction were based on the published sequence of murine Rab24 [ 24 ] . Kpn1 and BamH1 restriction sites were added to the 5' and 3' primers respectively, to facilitate cloning of the PCR product in frame with a 5' sequence cassette encoding a myc epitope (EQKLISEEDL), previously introduced into the pCMV5 expression vector [ 78 ] . The Rab24 cDNA was altered by overlap-extension PCR [ 79 ] , using appropriate mutator oligonucleotides and Pfu DNA polymerase (Stratagene Inc., La Jolla, CA), so as to introduce amino acid substitutions at D123 (D123I), T120 (T120A), S22 (S22N), S67 (S67Q), and a deletion of residues 127 through 130 (ΔEDRR).
As shown in fig 1athe correct integration of Neo at the hprt locus was predicted to delete a BamH1 restriction endonuclease cut-site from intron 1 and replace it with a BamH1 site that is unique to the mutant hprt allele.
PCR products were restriction digested with BamH1 (exon 1) or Not1 (exon 2), purified from an agarose gel by Qiaquick extraction (Qiagen), and simultaneously subcloned into a BamH1/Not1 digested pcDNA3 (Invitrogen) vector to create a directional clone of rabbit MYOC exons 1 and 2 spliced together (pcDNA3.RbMYOCex1-2).
Human wild type p53 or p53 bearing mutations in serines 33 or 37 were inserted into the BamH1 site of the pcDNA3.
After digestion with Nde I and BamH I the coding sequence was subcloned to the vector pET-11a (Stratagene) for gene expression experiments.