Notes: Total RNA was isolated from uninfected Vero cells and Vero cells infected with either Simkania negevensis Z^T or Chlamydia trachmatis using the SV Total RNA Isolation System. The isolated RNA was used in RT-PCR amplifications to determine the size of the unspliced intron form of the 23S rRNA to differentiate between a group I intron where the rRNA intron is spliced with religation and an intervening segment where the rRNA is fragmented to functionally remove the intervening segments. The isolated RNAs and Promega's RNA Markers were separated by electrophoresis to judge RNA integrity. (2302)

Notes: The RNA Markers, 0.28-6.58kb, were used for size calibration on RNA gels. (0409)

Notes: RNA probes were produced in the presence of the RNasin^® Ribonuclease Inhibitor. The probes were used in Northern blotting and the sizes of the reactive bands were determined in comparison to the RNA Markers, 0.28-6.58kb. All reagents for RT-PCR (Taq DNA Polymerase; MMLV Reverse Transcriptase; dNTPs and RNasin^® Ribonuclease Inhibitor) were obtained from Promega. (1485)

Notes: Tryptic peptides, produced with Sequencing Grade Modified Trypsin, of the purified lipase were used to generate primers for RT-PCR. The largest amplimer was purified and used to screen a lambda gt11 library. The isolated 303 amino acid clone and site-specific mutants were put into the pCI-neo Mammalian Expression Vector and expressed in COS cells. The transiently expressed proteins were assayed for esterase and lipase activity. (0960)