Lambda Exonuclease

Applications

  • Single-strand conformation polymorphism (SSCP) analysis of PCR products.1,2
  • Generation of ssDNA sequencing template from PCR products.

Lambda Exonuclease is a highly processive 5´→3´ exodeoxyribonuclease that selectively digests the 5´ phosphorylated strand of dsDNA. The preferred substrate is blunt-ended, 5´-phosphorylated dsDNA. The enzyme has reduced activity against nicked DNA and against ssDNA and gapped DNA.3

Unit Definition: One unit of Lambda Exonuclease will produce 10 nmol of acid-soluble deoxyribonucleotides from dsDNA template in 30 minutes at 37°C in 1X Lambda Exonuclease Reaction Buffer.

Quality Control: Lambda Exonuclease is function-tested to ensure complete and preferential degradation of PCR products produced using 5´-phosphorylated primers. PCR products made using primers containing 5´-OH ends are not digested. Lambda Exonuclease is tested to be free of contaminating endonuclease activities.

References

  1. Schwieger, F. and Tebbe, C.C. (1998) App. and Environ. Microb. 64, 4870.
  2. Schwieger, F. and Tebbe, C.C. (2000) App. and Environ. Microb. 66, 3556.
  3. Mitsis, P.G. and Kwagh, J.G. (1999) Nucleic Acids Res. 27, 3057.
Figure. 1 Lambda Exonuclease selectively digests the strand of a PCR product produced using a PCR primer with a 5´-phosphate. The resulting single-stranded PCR product can be used for SSCP analysis or sequencing.