The methods are also useful for subcloning large DNA fragments without the need for restriction enyzmes.

The present invention relates to improved methods of introducing site-directed mutations into circular DNA without the need for subcloning.

The present invention is directed to methods and compositions for DNA subcloning using bacterial recombinase-mediated homologous recombination.

An Nde1 site in the promoter provides an in-frame start methionine and a standard polylinker is available for ease of subcloning.

Subcloning the cDNA sequences into vectors and the preparation of cells stably transformed with the vectors are also disclosed.

Also disclosed are rapid subcloning methods employing the vectors and enzymes disclosed herein and kits for use in such methods.

This method eliminates the inefficient sub-cloning steps and the need for costly primer sets required by many prior methods.

The vaccines were constructed by subcloning cDNA representing the medium (M) (encoding the G1 and G2 glycoproteins) into the DNA expression vector pWRG7077.

Digital Bisulfite Genomic DNA Sequencing methods are also provided for high-resolution DNA methylation information without subcloning.

The bacterium is acquired by subcloning methionine enzyme genes in pSVSPORT particles, then by using the particles to transform attenuated Salmonella typhimurium VNP20009.

The method avoids the need for synthesizing long strands of nucleic with a DNA synthesizer and avoids the need for inefficient steps such as ligation and subcloning.

The present invention provides compositions, including vectors, and methods for the rapid subcloning of nucleic acid sequences in vivo and in vitro.

Micro-scale methods are applied in producing, maintaining, replicating, screening, manipulating and sub-cloning cell libraries.

Single-stranded RNA probes derived from the DNA clone after subcloning into an in vitro transcription vector were also used to show that the Norwalk virus contains an ssRNA genome of about 8 kb in size.

After sub-cloning, each segment can be used as a probe to identify cloned DNA sequences that, at the chromosome level, are roughly 200 kb apart.