Jacobs, A.T. and Ignarro, L.J.
Notes: Promega T4 Polynucleotide Kinase was used to end-label an oligo representing a NF-kB binding sequence element with [γ-32P] ATP (7,000 Ci/mmol). Specific primers to IFN-β and IκB-α messenger RNA were used in RT-PCR to generate products for cloning into the pGEM®-T Easy Vector. The resulting plasmids were linearized with Nco I and Spe I, respectively, and used as templates for in vitro transcription using the Riboprobe® System to generate probes for use in an RNase protection assay. The antisense probes were labeled with [α-32P]CTP (800 Ci/mmol) in the Riboprobe® reactions. RNase ONE™ Ribonuclease was also used in this study. (3197)