ssDNA

The Long ssDNA Preparation Kits (LsODN Preparation Kits) give a simple and easy method for generation of a long single stranded DNA (within 10,000 bases). A long ssDNA prepared by this method has defined sequence and length because it does not include inside mutation and terminal deletion caused by PCR, exonuclease side reaction, not high-fidelity reverse transcriptase reaction or not high-fidelity synthetic oligonucleotides. The procedure of this method is almost the same as the method to obtain dsDNA fragments. The DNA of interest is cloned into a plasmid. The resulting plasmid harboring the DNA is digested with a pair of two nicking endonucleases or a combination of a nicking endonuclease and a restriction enzyme. The nicked plasmid is denatured by mixing with Denaturing Gel-Loading Buffer and then subjected to agarose gel electrophoresis. The band corresponding to a long ssDNA is excised and extracted with commercially available kits.

1. The DNA of interest is cloned into a plasmid between the two Nicking enzyme sites, or between the Nicking enzyme site and the restriction enzyme site.

2. The resulting plasmid harboring the DNA is digested with a pair of two nicking endonucleases or a combination of a nicking endonuclease and a restriction enzyme.

3. The nicked plasmid is denatured and then subjected to agarose gel electrophoresis.

4. The band corresponding to a long ssDNA is excised and extracted.

Long ssDNA Gel Extraction Kit has been optimized for extraction of high-quality Long ssDNA from agarose gel.
There are two lineups according to ssDNA length.