商品编号


EG-1010



品牌


Excellgen



公司


Excellgen, Inc.



公司分类


DNA Modifying Enzymes



Size

20,000 Units

商品信息



Description


Exonuclease I cleaves single-stranded DNA in the
3' to 5'
direction, releasing 5'-mono/di-nucleotides and leaving double-stranded DNA molecules and the 5'-terminus intact. The enzyme is processive though digestion is inhibited by the presence of a 3'-terminal phosphate. This enzyme does not cleave DNA strands with terminal 3'-OH groups blocked by phosphoryl or acetyl groups




Applications



Primer removal from PCR mixtures:

prior to PCR product sequencing (2)

for one-tube "megaprimer" PCR mutagenesis (3)



Removal of single-stranded DNA containing a 3'-hydroxyl terminus from nucleic acid mixtures.

Assay for the presence of single-stranded DNA with a 3'-hydroxyl terminus (4)

In the prescence of lam
BD
a exonuclease, Exo I is effective in removing linear DNA from plasmid preparations.





Source

An
E. coli
strain that carries the cloned
E coli

sbcB
gene



Unit Definition


One unit of the enzyme catalyzes the release of 10 nmol of acid soluble nucleotides in 30 min at 37°C.




Molecular Weight

54.5 kDa monomer.



Components



Endonuclease I:
20,000 U/ml in 10 mM Tris-HCl, pH 7.5 @ 25°C, 100 mM NaCl, 1 mM DTT, 0.5 mM EDTA, 50% glycerol

10X Reaction Buffer
:
670 mM Glycine-KOH,
pH 9.5 @ 25°C, 67 mM MgCl
2
,
10 mM DTT.





Quality Control

The absence of other endo-, exodeoxyribonucleases, and ribonucleases confirmed by appropriate quality tests.



Storage Condition


-20 °C




References




I. R. Lehman, A. L. Nussbaum, The deoxyribonucleases of
Escherichia coli
. V. On the specificity of exonuclease I (phosphodiesterase).
J. Biol. Chem.

239
, 2628-2636 (1964).



E. Werle
et al.
, Convenient single-step, one tube purification of PCR products for direct sequencing.
Nucleic Acids Res.

22
, 4354-4355 (1994).



S. Nabavi, R. N. Nazar, Simplified one-tube “megaprimer” polymerase chain reaction mutagenesis.
Anal. Biochem.

2
, 346-348 (2005).



J. Rosamond
et al.
, Modulation of the action of the recBC enzyme of
Escherichia coli
K-12 by Ca
2+
.
J. Biol. Chem.

254
, 8646-8652 (1979).



Y. Sasaki, D. Miyoshi, N.Sugimoto, Regulation of DNA nucleases by molecular crowding.
Nucleic Acids Res.

35
, 4086-4093 (2007).