DNAse I

DNase I (Deoxyribonuclease I) bụ endodeoxyribonuclease nke nwere ike igbari DNA otu ma ọ bụ okpukpu abụọ.Ọ na-amata ma kewaa njikọ phosphodiester iji mepụta monodeoxynucleotides ma ọ bụ oligodeoxynucleotides nke nwere otu ma ọ bụ okpukpu abụọ nwere otu phosphate na 5'-terminal na hydroxyl na 3'-terminal.Ọrụ nke DNAse I dabere na Ca2+ ma enwere ike ịgbalite ya site na ion metal divalent dị ka Mn2+ na Zn2+.5mM Ca2+ na-echebe enzyme site na hydrolysis.N'ebe Mg2+ nọ, enzyme ahụ nwere ike ịmata ma kewaa saịtị ọ bụla na eriri DNA ọ bụla.N'ebe Mn2+ nọ, enwere ike ịmata eriri abụọ nke DNA n'otu oge wee kewaa ya n'ihe fọrọ nke nta ka ọ bụrụ otu saịtị ahụ iji mepụta iberibe DNA njedebe dị larịị ma ọ bụ iberibe DNA nke nnyapade nwere 1-2 nucleotides na-apụta.

Ngwongwo ngwaahịa

Bovine Pancreas DNAE gosipụtara m na usoro okwu yist ma sachaa ya.

Cndị ọchịchị

Ụgbọ njem na Nchekwa

1. Nchekwa Stability: - 15 ℃ ~ -25 ℃ maka nchekwa;

2.Transport Stability: Ụgbọ njem n'okpuru ngwugwu ice;

3. Enyere na: 10 mM Tris-HCl, 2 mM CaCl₂, 50% glycerol, pH 7.6 na 25 ℃.

Nkọwa nkeji

A kọwapụtara otu nkeji dị ka ọnụọgụ enzyme nke ga-eweda 1 μg nke pBR322 DNA kpamkpam n'ime nkeji iri na 37 Celsius.

Njikwa ogo

RNase:5U nke DNAse I nwere 1.6 μg MS2 RNA maka awa 4 na 37 ℃ anaghị eweta mbibi dịka agarose gel electrophoresis siri kpebie.

Ọrịa nje Endotoxin:LAL-ule, dị ka Chinese Pharmacopoeia IV 2020 mbipụta, gel ókè ule usoro, izugbe iwu (1143).Ọdịnaya endotoxin nke nje bacteria kwesịrị ịbụ ≤10 EU/mg.

Ntuziaka maka ojiji

1. Kwadebe ngwọta mmeghachi omume na tube na-enweghị RNase dị ka akụkụ ndị edepụtara n'okpuru:

2.37 ℃ maka nkeji 15;

3.Tinye ihe nkwụsị nkwụsị iji kwụsị mmeghachi omume ahụ, na-ekpo ọkụ na 65 ℃ maka nkeji 10 iji mebie DNAse I. Enwere ike iji ihe nlele ahụ mee ihe ozugbo maka nyocha ntụgharị ọzọ.

Ihe ndetu

1.Jiri 1U DNAse I kwa μg nke RNA, ma ọ bụ 1U DNAse I maka ihe na-erughị 1μg nke RNA.

Ekwesịrị ịgbakwunye 2.EDTA na ntinye ikpeazụ nke 5 mM iji chebe RNA ka ọ ghara ịda mbà n'oge mmechi enzyme.