RNArticles: RNA Amplification Used to Develop a New In Vitro HCMV Assay ( correlate their e...)

RNArticles: RNA Amplification Used to Develop a New In Vitro HCMV Assay

correlate their expression with in vivo infected bone marrow cells.

HCMV is a ubiquitous herpes virus found in 80% of the population where it is maintained in a latent form. Infection of healthy normal individuals by HCMV is usually harmless, but reactivation of the latent virus in immune compromised individuals can be life threatening. The latent viral replication can be activated in a subset of bone marrow cells. Unfortunately, it is currently not possible to work with these cells in culture nor has there been an in vitro model for the study of the latent virus (3).

Recently, Goodrum et al. (2) used microarray analyses to establish the role of infected primary CD34+ haemotopoeitic progenitors grown above a murine stromal AFT024 monolayer as a potential in vitro model system to study HCMV latency. For this study, HCMV cDNA arrays comprised of PCR products from 222 known ORF's for the AD169 and Toledo strains of HCMV (2,4) were tested with cDNA probes made from primary human fibroblasts or human CD34+ progenitor cells infected in vitro with HCMV. RNA was isolated from 10,000-20,000 infected CD34+ cells and then amplified using Ambion's MessageAmp aRNA Amplification Kit. For the human fibroblasts, total RNA was isolated and DNase I treated with Ambion's DNA-free Kit. Poly(A) RNA selection was carried out using the Ambion '"/>

Source:

Page: All 1 2 3 4
Related biology technology :

1. RNArticles: Keeping RNA Intact Prior to RNA Isolation - RNAlater
2. prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification
3. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
4. Greater Amplification Specificity with New Hot Start PCR Enzyme
5. Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase
6. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
7. Improve Amplification Specificity with Hot Start PCR Enzyme
8. Amplification of the amoA Gene of Nitrosococcus oceani Bacteria Using Eppendorf MasterTaq
9. Amplification of Mouse cDNAs for Microarrays Using the Eppendorf MasterTaq Kit
10. Optimizing the Direct Amplification of Missing cDNA 5 Ends Using the Eppendorf Mastercycler gradient
11. Optimizing DNA Amplification Protocols using the Eppendorf Mastercycler

Post Your Comments:

( correlate their e...)RNArticles: RNA Amplification Used to Develop a New In Vitro HCMV Assay