The Exonuclease I (ExoI) degrades single-stranded DNA in a 3'=>5' direction, releasing deoxyribonucleoside 5'-monophosphates in a stepwise manner and leaving 5'-terminal dinucleotides intact. It does not cleave DNA strands with terminal 3'-OH groups blocked by phosphoryl or acetyl groups.
Source:E. coli cells with a cloned E. coli sbcB gene.
Concentration:~20u/ul
Unit Definition:One unit of E9007-85C catalyzes the release of 10 nano-moles of acid soluble nucleotides in 30 min at 37ºC.
Form:Supplied as a liquid in 20mM Tris-HCl, pH 7.5, 0.1mM EDTA, 1mM DTT and 50% glycerol.
Supplied with:E9007-85C1: 10X Reaction Buffer: Supplied as a liquid in 670mM glycine-KOH, pH 9.5 at 25°C, 67mM MgCl2, 10mM DTT.
Applications:Suitable for use in the purification of PCR products, removal of single-stranded DNA containing a 3'-hydroxyl terminus from nucleic acid mixtures and assay for the presence of single-stranded DNA with a 3'-hydroxyl terminus. Other applications not tested.
Protocol for Elimination of Oligonucleotides and dNTPs from PCT mixtures:1. Remove a 5ul aliquot from a reaction mixture
2. Add 10u of E9007-85C and 1u of Shrimp Alkaline Phosphatase
3. Mix and incubate at 37ºC for 15 min
4. Inactivate the enzymes by heating at 85ºC for 15 min
*Note: After inactivation, the aliquot can be used directly for DNA sequencing. The Exonuclease I is not suitable for removing 3’-overhangs of dsDNA.
Storage and Stability:May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size：4000U
Source Antigen：E. coli cells with a cloned E. coli sbcB gene.
