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Example sentences for: stratagene
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The resulting cDNA inserts were purified by agarose gel electrophoresis, digested with Eco R I and cloned into pBluescript KS- (Stratagene, La Jolla, CA).
mRNA was extracted from murine tissues (BALB/c mice: Id wound of controls, 1d wound of mice treated with 0.5 mg Dexamethason per kg bodyweight twice a day for 5 days before wounding) according to standard procedures [ 30 ] . Methylated cDNA was synthesized from an anchored Xho I-oligo-dT primer ((GA) 10 ACTAGTCTCGAGT 16 VN) that secured a defined start of the cDNA synthesis at the cDNA-polyA border using the Stratagene cDNA synthesis kit according to the instructions of the supplier.
Mutazyme (Stratagene, La Jolla, CA) was used to amplify PcrG for cloning into pBAD18 [ 33 ] . The ligation reaction was electroporated into lcrG Y. pestis (which is temperature sensitive due to its Ca 2+-blind phenotype) and the cells were plated on TMH medium [ 29 ] containing arabinose, Ca 2+, kanamycin and carbenicillin.
Following isolation of pure plaques, the insert DNAs were prepared by PCR methods with the following conditions: 100 μl reactions containing 1X Pfu buffer (20 mM Tris-Cl, pH 8.75, 10 mM KCl, 2 mM MgSO 4 , 10 mM (NH 4 ) 2 SO 4 , 0.1% v/v Triton X-100, 100 μg/ml bovine serum albumin), 1 μM λgt11 forward and reverse primers (Promega, Madison, WI), 0.2 mM deoxyribonucleotides, and 5 Units of recombinant Pfu DNA Polymerase (Stratagene, La Jolla, CA).
These cDNAs were gel-purified and used to produce 32P-labeled cDNA probes (Prime-It kit, Stratagene).
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