Exo-Resistant Random Primer

Exo-Resistant Random Primer (5´-3´)

Protects from 3´→ 5´ exonuclease degradation and ensures efficient priming of DNA synthesis. For Research Use Only.


Description
MCLAB´s Exo-Resistant Random Primer is a mixture of single-stranded random oligonucleotides. It can be used in many applications such as highly efficient random priming of various DNA synthesis reactions. The primer in this prodcut has two 3´-terminal phosphorothioate (PTO) modifications that are resistant to the 3´→ 5´ exonuclease activity of proofreading DNA polymerases (1), like Klenow Fragment and phi29 DNA Polymerase. It also has 5´- and 3´-hydroxyl ends.

The product is supplied as a ready-to-use, 20X concentrated aqueous solution.

Applications

• Strand displacement amplification of genomic DNA (2), plasmids and phage DNA (3).
• DNA labeling by random priming (4-6).

• 500 µM (1.1 µg/µl)

• Functionally tested for the efficient priming of DNA synthesis using phi29 DNA Polymerase.

1. Skerra, A., Phosphorothioate primers improve the amplification of DNA sequences by DNA polymerases with proofreading activity, Nucleic Acids Res., 20, 3551-3554, 1992.
2. Dean, F.B., et al., Comprehensive human genome amplification using multiple displacement amplification, Proc. Natl. Acad. Sci., 99, 5261-5266, 2002.
3. Dean, F.B., et al., Rapid amplification of plasmid and phage DNA using phi29 DNA polymerase and multiply-primed rolling circle amplification, Genome Res., 11, 1095-1099, 2001.
4. Feinberg, A.P. and Vogelstein, B., A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity, Anal. Biochem., 132, 6-13, 1983.
5. Feinberg, A.P. and Vogelstein, B., A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity, Addendum, Anal. Biochem., 137, 266-267, 1984.
6. Mackey, J., et al., Use of random primer extension for concurrent amplification and nonradioactive labeling of nucleic acids, Anal. Biochem., 212, 428-435, 1993.