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 in 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 primers ^(4-6).

Concentration:
500 µM (1.1 µg/µl)

Quality Control:
Functionally tested for the efficient priming of DNA synthesis using phi29 DNA Polymerase.

References:
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.