DNA into a cloning vehicle

This is a random process but among the products will be recircularized vehicles into which has been ligated the DNA sequence of interest. As discussed earlier an insertion into an antibiotic resistance gene provides a simple selection procedure for isolating the recombinant plasmids. Many variations of this basic 

In some circumstances the aim of the experiment will be to clone a DNA sequence which lacks cohesive ends. Such molecules may, for example, be the cDNA products from the reverse transcription of particular mRNA species and this is currently the most widely used procedure for sequencing eukaryotic mRNAs. Under controlled conditions the reverse transcriptase from Avianmyeloblas-tosis virus (AMV) will use a mRNA template to synthesize a full-length double-stranded DNA copy (cDNA) of the RNA sequence. The products of this reaction frequently lack precisely defined ends and it is necessary to trim such frayed or uneven termini before proceeding to the cloning step. Incubation with DNA polymerase (Klenow) or T4-polymerase (Section 4.2.2.) will yield molecules with clean blunt-ended termini which can be 

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