Application of mRNA reverse transcription amplification of cDNA (RT-PCR)
RT-PCR can be used to: (1) construct large-capacity cDNA libraries; (2) identify mutations and polymorphisms in transcribed sequences; (3) measure the intensity
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Ribonuclease protection (mapping of RNA with ribonuclease and radiolabeled RNA probes)
Ribonuclease protection analysis is used to measure the abundance of specific mRNAs and to map their topological heterogeneity. This method involves hybridizati
Application of mixed oligonucleotide primer-guided cDNA amplification (MOPAC)
The best way to clone a protein for which only part of the sequence is known is to design an oligonucleotide using the known amino acid sequence and use this ol
Introduction of restriction endonuclease sites at the end of amplified DNA products by PCR amplification
The source of this experiment is "Guide to Molecular Cloning Experiments, Third Edition", translated by Huang Peitang et al.
Analysis of RNA by primer extension method
Primer extension is primarily used for mRNA 5' end mapping. poly (A) + RNA is first hybridized to an excess of a 5' end-labeled single-stranded oligonucleotide
Rapid amplification of cDNA 5' ends (5'-RACE)
There is no greater failure in molecular cloning than the isolation of a cDNA clone that lacks the characteristic structure of the sequence at the 5' end of the
Labeling the 3' protruding end of double-stranded DNA with [α-32P] 3'deoxyadenosine 5'-triphosphate or [α-32P] dideoxyATP ends
The source of this experiment is "Guide to Molecular Cloning Experiments, Third Edition", translated by Huang Peitang et al.
Random priming method (radiolabeling of DNA using random oligonucleotide extensions in the presence of melted agarose)
This method can be used for radiolabeling of DNA recovered from low melting point agarose gels (Feinberg and Vogelstein 1983, 1984). This experiment is based on
Preparation of radiolabeled DNA probes using the polymerase chain reaction
The source of this experiment is "Guide to Molecular Cloning Experiments, Third Edition", translated by Huang Peitang et al.