What is CBI’s custom long single-stranded DNA (long ssDNA, lssDNA)?
We offer custom long single-stranded DNA (long ssDNA, lssDNA) (>200 bases). We directly create long ssDNA (lssDNA) fragments from large amounts of plasmid. This new method completely removes the need for PCR and reverse transcription. Hence, our long ssDNA fragments are featured with the precise sequence. In addition, our method has no limits to the gene length. Our ssDNA fragments are useful for CRISPR-mediated gene knock-in, in vitro transcription, SNP detection, antibody development, and other applications.
In contrast, chemical methods can only produce long ssDNA fragments up to a few hundred bases. As a result, the generation of a long ssDNA fragment often involves PCR or reverse transcription of an RNA intermediate. Such enzymatic methods, however, can cause mutations to ssDNA products. Additionally, the attainable length of long ssDNA fragments is still limited. Thus, our long single-stranded DNA oligo synthesis method is more favorable for the creation of high-quality long ssDNA fragments.
•Any length ssDNA oligos
•No DNA bases damage
•No double-stranded DNA (dsDNA) fragment contamination
•Small to large scale production
•Oligo synthesis and assembly
•Cloning and sequence verification of the full-length gene
•Expansion of verified clone
•Production of long ssDNA
•ssDNA fragment dry powder
•Certificate of Analysis (COA)
For gene knock-in, long ssDNA oligos (lssDNA) are more favorable than dsDNA donors
In gene knock-in experiments, both long ssDNA and dsDNA donors are efficient HDR templates for inserting long genes. However, dsDNA donors are very likely to insert at off-target genomic locations. Compared to dsDNA HDR repair templates, long ssDNA fragments (lssDNA) can drastically reduce the tendency to randomly insert into the genome. As a result, ssDNA repair templates can improve the precision of insertion. Additionally, transfection of a long ssDNA (lssDNA) fragment is far less toxic to cells than a dsDNA template. Furthermore, long ssDNA (lssDNA) has no expression from non-integrated templates. Thus, it is easier to identify and isolate correctly edited clones. Our high-quality custom long ssDNA fragments (lssDNA) are very useful for this application.
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