69202  Strandase™ ssDNA Preparation Kit

Rapid conversion of double-stranded PCR products to single-stranded DNA

The Strandase ssDNA Preparation Kit is designed for convenient production of single-stranded DNA templates suitable for sequencing or other applications. The kit contains Strandase λ Exonuclease, which digests one strand of duplex DNA from a 5′-phosphorylated end, releasing 5′-phosphomononucleotides (Little 1967). Suitable DNA templates can be generated by PCR in which one of the primers contains a 5′-terminal phosphate. In templates containing one phosphorylated primer and one nonphosphorylated primer, the strand with the phosphorylated primer is selectively degraded.

In the Strandase procedure, PCR products are first precipitated with sodium acetate and isopropanol to concentrate the DNA and remove primers and other reaction components. The DNA is then digested with Strandase λ Exonuclease, heated to inactivate the enzyme, and added directly to a sequencing reaction. The Strandase method produces concentrated, purified ssDNA that can be used for downstream applications.

Vector-specific primers and T4 polynucleotide kinase with the appropriate buffer are available for convenient preparation of phosphorylated primers.