DNAse I (N'efu)(5u/ul)
Mpempe nwamba: HC4007A
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 DNAse I dabere na Ca2+enwere ike ịgbalite ya site na ion ọla divalent dị ka Mn2+na Zn2+.5 mM ca2+na-echebe enzyme site na hydrolysis.N'ihu Mg2+, enzyme ahụ nwere ike ịmata ma kewaa saịtị ọ bụla na eriri DNA ọ bụla.N'ihu Mn2+, 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 na-adọrọ adọrọ nke nwere 1-2 nucleotides na-apụta.
Ngwa
| Aha | 0.1KU | 1KU | 5 KU | 50 KU |
| DNAse I, RNase enweghị | 20μL | 200μL | 1 ml | 10 ml |
| 10×DNase I nchekwa | 1 ml | 1 ml | 5 × 1 ml | 5 × 10 ml |
Ọnọdụ nchekwa
-25 ℃ ~ -15 ℃ maka nchekwa;Ụgbọ njem n'okpuru ngwugwu ice.
Ntuziaka
1. Kwadebe ngwọta mmeghachi omume na tube na-enweghị RNase dị ka oke nke edepụtara n'okpuru:
| Akụkụ | Olu |
| RNA | X µg |
| 10 × DNAse I nchekwa | 1μl |
| DNAse I, RNase n'efu (5U/μL) | 1 U kwa µg RNA① |
| ddH2O | Ihe ruru 10 μl |
Mara: ① Gbakọọ olu DNAse I nke kwesịrị ịgbakwunye dabere na ọnụọgụ RNA.
2. 37 ℃ maka nkeji iri na ise;
3. Tinye 0.5M EDTA na njedebe ikpeazụ nke 2.5mM ~ 5mM, na okpomọkụ na 65 ℃ maka nkeji 10 iji kwụsị mmeghachi omume ahụ.Enwere ike iji ihe nlele ahụ ozugbo maka mmeghachi omume na-esote dị ka ntụgharị ntụgharịnnwale.
Nkọwa nkeji
A kọwapụtara otu nkeji dị ka ọnụọgụ enzyme nke ga-eweda 1µg nke pBR322 kpamkpamDNA na nkeji 10 na 37 ℃.
Njikwa ogo
RNase:5U nke DNAse I nwere 1.6µg MS2 RNA maka awa 4 na 37 ℃ anaghị eweta mbibi dịkaA na-ekpebi site na agarose gel electrophoresis.
Ihe ndetu
1. Biko kwadebe 0.5MEDTA n'onwe gị.
2. Jiri 1U DNAse I kwa µg nke RNA.Agbanyeghị, ọ bụrụ na RNA erughị 1µg, biko jiri 1U DNAse I.
3. Biko tinye enzyme na ice n'oge ọrụ.
