Lambda Exonuclease

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Last modified: 2008-07-15 14:57:15

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

Unit Definition:

One unit will produce 10nmoles of acid soluble deoxyribonucleotides from double-stranded DNA 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 product produced using 5'-phosphorylated primers. PCR product made using primers containing 5'-OH ends is not digested. Lambda Exonuclease is tested to be free of contaminating endonuclease activities.

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.

Last modified: 2008-05-14 16:46:46

Applications

• SSCP (single-strand conformation polymorphism) analysis of PCR product.
• Generate single-stranded DNA sequencing template from PCR product.

Last modified: 2008-05-14 16:46:46

Protocols for: Lambda Exonuclease

(catalogue number LE035H / LE032K)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-14 16:46:46

References

1. Mitsis, P.G. and Kwagh, J.G. (1999) Nucl. Acids. Res. 27 (15), 3057.
2. Schwieger, F. and Tebbe, C.C. (1998) App. and Environ. Microb. 64 (12), 4870.
3. Schwieger, F. and Tebbe, C.C. (2000) App. and Environ. Microb. 66 (8), 3556.

Last modified: 2008-05-14 16:46:46