E-SMDB-2961 - Transcription profiling of background variability of HFFs
Submitted on 7 November 2005, released on 7 November 2005, last updated on 4 June 2014
Background variability was determined by hybridizing Cy3- and Cy5-cDNA from the same source (mock infected HFFs) to arrays (HE series). Polyadenylated RNA from 107 confluent HFs was purified by using Oligotex (QIAGEN, Valencia, Calif.) from Trizol (Invitrogen)-extracted total RNA. This RNA was prepared, reverse transcribed, labeled with Cy3-dUTP or Cy5-dUTP (Amersham, Little Chalfont, Buckinghamshire, United Kingdom) by random primed synthesis with DNA pol I Klenow (Amersham Life Science, Inc., Cleveland, Ohio), and hybridized to spotted, human cDNA microarrays as previously described. Sequence-verified human cDNA microarrays (HE and HG series, 31,000 spots; HD51 series, 17,000 spots) were produced at Stanford. Images were collected by using a GenePix 4000B microarray scanner, manually flagged to eliminate poor spots and analyzed by GenePix Pro 2.0 (Axon, Union City, Calif.).
transcription profiling by array, quality control testing
Major human cytomegalovirus structural protein pp65 (ppUL83) prevents interferon response factor 3 activation in the interferon response. Abate DA, Watanabe S, Mocarski ES. J Virol 78(20):10995-1006 (2004), Europe PMC 15452220