AFLP ‐Based Transcript Profiling
| Autor: | Pieter Vos, Patrick Stanssens |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Differential display
DNA Complementary Transcription Genetic cDNA library RNA DNA Templates Genetic Computational biology Biology Molecular biology Restriction enzyme Rapid amplification of cDNA ends Transcription (biology) Complementary DNA Indicators and Reagents Amplified fragment length polymorphism RNA Messenger Amplified Fragment Length Polymorphism Analysis Deoxyribonucleases Type II Site-Specific Molecular Biology |
| Zdroj: | Current Protocols in Molecular Biology |
| ISSN: | 1934-3647 1934-3639 |
| DOI: | 10.1002/0471142727.mb25b05s57 |
| Popis: | This unit presents an alternative to differential display that allows the quantification of transcripts, based on AFLP-fingerprinting of double-stranded cDNA. The protocol described includes the following steps: the isolation of poly(A)+ RNA from total RNA, the synthesis of double-stranded cDNA, the preparation of template fragments by digestion of the cDNA library with a combination of two restriction enzymes and the ligation of adaptors to the fragment ends, the selective amplification of specific subsets of fragments, and the electrophoretic analysis of these amplification products on standard denaturing polyacrylamide gels. The transcript profiles obtained by this technique are a reliable and efficient tool to identify differentially expressed mRNAs. This unit presents an alternative to differential display that allows the quantification of transcripts, based on AFLP-finger his unit presents an alternative to differential display that allows the quantification of transcripts, based on AFLP-finger. |
| Databáze: | OpenAIRE |
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