Improved Long ssDNA preparation kits

Specially designed purification kit is now included!
Long single strand DNA (ssDNA) preparation kit
For effective knock-in by genome editing

The Long ssDNA Preparation Kits (LssDNA Preparation Kits) provide a simple and easy method for generating a long ssDNA (<1,500 base or < 3,000 base).
A long ssDNA prepared by this kit has a defined sequence and length as it does not include internal mutation and terminal deletion caused by PCR, exonuclease side reaction, low-fidelity reverse transcriptase reaction or low-fidelity synthetic oligonucleotides.

The procedure 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 by a pair of two nicking endonucleases or a combination of a nicking endonuclease and a restriction enzyme.
The nicked plasmid is denatured by Denaturing Gel-Loading Buffer and then subjected to agarose gel electrophoresis.
The band corresponding to a long ssDNA is excised and extracted with our newly developed extraction kits.

(1) The DNA of interest is cloned into a plasmid using a pair of two nicking endonuclease sites or a combination of a nicking endonuclease site and a 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.

Fig.1 Agarose gel electrophoresis of purified long ssDNAs.
1.2% agarose gel in 1x TAE. 5.5cm x 6.0 cm.
Lane 1: pLSODN-1 (1.5 kb Fragment), double-nicked and denatured 800 ng
Lane 2: a long ssDNA purified from pLSODN-1 (1.5 kb Fragment) 400 ng
Lane 3: pLSODN-3 (3 kb Fragment), double-nicked and denatured 800 ng
Lane 4: a long ssDNA purified from pLSODN-3 (3 kb Fragment) 400 ng

DS615 Long ssDNA Preparation Kit for 1.5 kb (LsODN Preparation Kit)
• pLSODN-1 10 μg (0.5 μg/μl)
• pLSODN-2D 10 μg (0.5 μg/μl)
• Denaturing Gel-Loading Buffer 1 ml (100 loadings)
• Nicked pLSODN-1 (1.5 kb Fragment)
• Long ssDNA Gel Extraction Kit for 3kb (25 preps)

DS625 Long ssDNA Preparation Kit for 3.0 kb (LsODN Preparation Kit)
• pLSODN-3 10 μg (0.5 μg/μl )
• pLSODN-4D 10 μg (0.5 μg/μl )
• Denaturing Gel-Loading Buffer 1 ml (100 loadings)
• Nicked pLSODN-3 (3.0 kb Fragment)
• Long ssDNA Gel Extraction Kit for 3kb (25 preps)

(1) LssDNA kit has two price systems now (Academic or Industrial).
Please contact your local distributor about price.
(2) Letter of Assurance (LoA) is required upon ordering.

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