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1Division of Immunology and Allergy,Lausanne University Hospital, 2Department of Fundamental Neuroscience,University of Lausanne, 3Department of Biochemistry,Erasmus Medical Center, 4Theoretical Biology and Biophysics Group,Los Alamos National Laboratories, 5Department of Biochemistry,University of Lausanne
Chapters
- 00:04Title
- 01:14Reverse Transcription and RNA Isolation
- 02:49Affinity Purification of ASP Biotinylated cDNA
- 04:29Standard PCR
- 05:10Results: ASP RNA Detection in CD4+ Cells from HIV Patients
- 06:56Conclusion
RNA hairpins and loops can function as primers for reverse transcription (RT) in absence of sequence-specific primers, interfering with the study of overlapping antisense transcripts. We have developed a technique able to identify strand-specific RNA, and we have used it to study HIV-1 antisense protein ASP.
Tags
Keywords: HIV-1Antisense Protein (ASP)RNA TranscriptsStrand-specific RT-PCRSense And Antisense SequencesBiotinylated PrimersRNA DenaturationAnti-sense CDNAsOverlapping GenesQuantificationDNase TreatmentReverse TranscriptionEndogenous RT ControlsReaction MixtureMagnetic BeadsWashing And Binding Buffer
