Nonradioactive ribonuclease protection analysis using digoxygenine labeling and chemiluminescent detection
- 1 January 1992
- journal article
- research article
- Published by Wiley in Electrophoresis
- Vol. 13 (1) , 637-638
- https://doi.org/10.1002/elps.11501301130
Abstract
A sensitive nonradioactive ribunouclease protection assay is described which we have used to study c-myc gene transcription and promoter usage in GLC4, a human small cell lung carcinoma cell line with amplified gene. For in vitro transcription we used digoxygenine (DIG)-rUTP instead of [α-32P]CTP or [α-32P]UTP and after polyacrylamide gel electrophoresis the protected probes were transferred to a nylon membrane from Boehringer Mannheim using electroblotting. Subsequently the membrane was analyzed by chemiluminescent detection. Results were obtained after 1 h of exposure and were comparable with those using radioactivity.Keywords
This publication has 4 references indexed in Scilit:
- Detection of Mutations in Human Type I Collagen mRNA in Osteogenesis Imperfecta by Indirect RNase ProtectionJournal of Biological Chemistry, 1989
- [67] A molecular titration assay to measure transcript prevalence levelsPublished by Elsevier ,1987
- Recent Advances in the Development of Methods for Detecting Single-base Substitutions Associated with Human Genetic DiseasesCold Spring Harbor Symposia on Quantitative Biology, 1986
- A method to detect and characterize point mutations in transcribed genes: amplification and overexpression of the mutant c-Ki-ras allele in human tumor cells.Proceedings of the National Academy of Sciences, 1985