Words similar to digested
Example sentences for: digested
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Each of these PCR products was cloned individually into the Eco RV site of pBluescript KS+, and then restriction digested with Eco RI and Not I (first-set fragment) or Not I and Pst I (second-set fragment).
The resulting cDNA inserts were purified by agarose gel electrophoresis, digested with Eco R I and cloned into pBluescript KS- (Stratagene, La Jolla, CA).
Placental tissue was digested and trophoblast cells isolated and cultured as described previously [ 15 36 ] . Briefly, the cells collected after digestion were pooled, washed twice in DMEM containing 25 mM HEPES and 4500 mg/L glucose (DMEM-HG), diluted to 10 ml in DMEM-HG supplemented with heat inactivated 5% fetal bovine serum (FBS) and antibiotics (50 μg/ml gentamycin, 100 U/ml penicillin, and 100 μg/ml streptomycin), and viable large nucleated cells (95% in trypan blue exclusion test) were counted.
The adenovirus (AV) backbone for the aquaporin-1 (AQP1) sense and antisense AV constructs was a replication-deficient "first-generation" AV with deletions of the E1 and E3 genes [ 26 ] . This "empty" AV contains the cytomegalovirus (CMV) promoter and bovine growth hormone polyadenylation (bHG) site separated by a polylinker that was used to clone AQP1 DNA as described previously [ 27 ] . A recombinant AQP1 virus was constructed using a plasmid containing the coding sequence for AQP1, pCHIPev [ 28 ] . pCHIPev was digested and the AQP1 insert was subcloned into the shuttle vector pSKAC, creating pSKAC/AQP1.
The PCR products were digested with Eco RI and Sal I and ligated into the Eco RI and Sal I restriction sites of pGAD424 to produce constructs pGAD424/Fib-1C EGF#1-5 and pGAD424/Fib-1C EGF#5-COOH respectively.
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