Words similar to pbluescript
Example sentences for: pbluescript
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Each PCR product was gel-purified and cloned into the Eco RV site of pBluescript KS+ (Stratagene).
The rat VEGF probe is a 0.9 kb cDNA Pst I/Sma I insert cloned into a pBluescript II KS+ vector [ 33 ] . Prehybridization and hybridization were performed in 50% formamide, 5×SSC (20×SSC is 0.3 M sodium chloride, 0.3 < sodium citrate), 10× Denhardt's solution (50× Denhardt's solution is 2% Ficoll, 2% polyvinyl pyrrolidone, 2% Bovine Serum Albumin Factor V), 50 mM sodium phophate (pH 7.0), 1% sodium dodecyl sulfate (SDS), and 250 μg/ml salmon sperm DNA at 42°C.
cDNA templates were subcloned into the pBluescript KS-plasmid and completely sequenced from both strands.
Purified TOP1 was pre-incubated with GST or GST-BTBD2 for 20 min before addition of the DNA substrate, which was either an end-labeled 36 bp fragment containing a single strong TOP1 cleavage site [ 24 ] , or a 161 bp PvuII-HindIII fragment of pBluescript (Stratagene) [ 25 ] ). End labeling of the DNA substrates was as previously described [ 24 25 ] . Cleavage reactions were continued for 20 min before stopping with 0.5% SDS.
We digested previously isolated lambda genomic clones of the human FGF8 region [ 22 ] with restriction endonucleases (Promega, Madison, WI) and analyzed them by Southern blotting using a mouse Npm3 cDNA [ 21 ] probe, as previously described [ 20 ] . An 8.4-kb Xho I fragment from these clones that hybridized to the probe was subcloned into pBluescript (Stratagene) for analysis.