Words similar to spei

Example sentences for: spei

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To construct [ pMET3-CDC15, URA3 ] (pWS100) and [ pMET3-cdc15-2, URA3 ] (pWS109), the corresponding genes (from start codon to 300 bps downstream of the stop codon) were amplified by PCR from genomic DNA of wild type (RJD381) or cdc15-2 (RJD619) cells, and cloned into the SpeI and SacII sites of RDB620 (a derivative of pRS316 containing the MET3 promoter inserted between the HindIII and EcoRI sites).
The resulting double-stranded DNA was diluted 1:50 and amplified using primers SpeI.
The inserts were released by EcoRI and SpeI digestion and sized using agarose gel electrophoresis.
The wild type plasmid pTZtd1304 [ 17 ] contains a fully functional 265 nt derivative of the 1016 nt wild type intron [ 15 ] . This derivative also contains the substitutions U34A, which introduces a SpeI site, and U976G, which introduces an EcoRI site.
The PCR product was digested with SpeI (New England Biolabs, Beverly, MA; 20 units) and EcoRI (50 units) in buffer (50 mM NaCl, 100 mM Tris-HCl, pH 7.5, 10 mM MgCl 2 , 0.025% Triton X-100, 100 μg/ml BSA) at 37°C for 60 minutes, purified, and cloned into SpeI/EcoRI digested pTZtd1304.