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Example sentences for: piscataway

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cDNA was labeled with [ 32P] and purified using Nick columns (Amersham Pharmacia Biotech AB, Piscataway, NJ).
Quantitation of specific mRNAs by RT-PCR was done essentially as described [ 34 ] . Briefly, total RNA was isolated from trypsinized cell monolayers using the Quickprep Micro™ mRNA purification kit from Pharmacia Biotech (Piscataway, NJ).
5% SDS-polyacrylamide gels, and transferred to nitrocellulose membrane (Amersham Piscataway, NJ) via semi-dry transfer.
For cytokines assays, 1 × 10 6transduced cells were incubated on anti-CD3 coated plates for 72 h at 37°C/5% CO 2 . Culture supernatants were then harvested and an ELISA assay was performed to measure the production of IL-4, IL-10 and IFN-γ as previously described [ 30 ] . To assay proliferation, the transduced cells were plated in duplicate in microtiter plates (5 × 10 5/well) at a ratio of 1:5 with irradiated BALB/c splenocytes and the indicated doses of OVA peptide at 37°C/5% CO 2 . After 72 h, 50 μCi/ml [ 3H]thymidine (Amersham, Piscataway, NJ) was added to the well.
RNA samples (5 μg) were reverse transcribed using random primers at annealing temperatures of 65°C (You-Prime-the-First-Strand kit, Amersham, Piscataway, NJ).