제품 설명Antarctic Phosphatase는 Antarctic TAB5 균주의 alkaline phosphatase 유전자를 E. coli에서 발현한 재조합 단백질입니다. 이 효소는 DNA, RNA 또는 NTP의 3' 또는 5' 말단에서 인산기를 제거합니다. 유전자 클로닝 과정에서 벡터의 self-ligation 이 일어나는 것을 방지하여, 원하지 않는 부산물의 생성을 줄일 수 있습니다.
제품 특징
- Isolated from a recombinant source
- Reaction temperature: 37℃
- Heat inactivation: 80℃ for 2 min
제품 응용
- Removing 5´ phosphates from DNA, RNA, rNTPs and dNTPs.
- Prevention of recircularization of cloning vectors.
- Removal of dNTPs and pyrophosphate from PCR reactions.
제품 구성품
- Antarctic Phosphatase
- 10X Antarctic Phosphatase Buffer
- Sterile water
Unit 정의
One unit is defined as the amount of enzyme that will dephosphorylate 1 μg of pUC19 vector DNA cut with Hind III (5´ protruding ends), Hinc II (blunts ends) or Pst I (5´ recessed ends) in 30 minutes at 37°C. Dephosphorylation is defined as > 95% inhibition of recirculation in a self-ligation reaction and is measured by transformation into E. coli.
Storage buffer
10 mM Tris-HCl (pH 7.4), 1 mM MgCl
2, 0.01 mM ZnCl
2, 1 mM DTT, 50% Glycerol.
10X Antarctic phosphatase Buffer
500 mM Bis-Tris-Propane HCl, 10 mM MgCl2, 1 mM ZnCl2, pH 6.0
참고사항
Antarctic Phosphatase의 활성엔 0.1 mM Zn2+ 가 요구됩니다.
Figure 1. Removal of 5’phosphates from ss/dsDNA by Antartic Phosphatase
15 fmol of 5’ end-labeled ss/dsDNA substrates were incubated with 5 unit of Antarctic Phosphatase at 25℃. Reaction products were removed at indicated time period and analyzed on PAGE.
