DNAse I (N'efu) (2u/ul)
Mpempe nwamba: HC4007B
DNAse I bụ endonuclease nke nwere ike igbari DNA nwere eriri abụọ ma ọ bụ otu.Ọ nwere ike hydrolyze phosphodiester bond iji mepụta mono-na oligodeoxynucleotides nwere otu 5'-phosphate na otu 3'-OH.Oke pH kacha arụ ọrụ nke DNAse I bụ 7-8.Ọrụ DNAse I dabere na Ca2+enwere ike ịgbalite ya site na ion metal divalent dị ka CO2, Mn2+, Zn2+, wdg N'ebe Mg2+, DNA Enwere m ike ihichapụ saịtị ọ bụla nke DNA nwere eriri abụọ na-enweghị usoro;Mgbe ọ nọ n'ihu Mn2+, DNase M nwere ike ịkewaa DNA nke nwere eriri abụọ n'otu saịtị ahụ, na-akpụ isi na-adịghị mma ma ọ bụ nnyapade na 1-2 nucleotides na-apụta.Enwere ike iji ya maka nhazi nke ụdị RNA dị iche iche.
Ngwa
| Aha | 1KU | 5KU |
| DNAseI recombinant (RNase na-enweghị) | 500 μl | 5 × 500 μl |
| Ihe nchekwa DNAse I Reaction (10×) | 1 ml | 5 × 1 ml |
Ọnọdụ nchekwa
Ekwesịrị ịchekwa ngwaahịa a na -25 ~ -15 ℃ maka afọ 2.Biko zere ifriizi ugboro ugboro.
Ntuziaka
Tinyere na mwepụ nke DNA na nlele RNA maka naanị ntụaka.
1. Biko jiri tubes centrifuge na-enweghị RNase na ndụmọdụ pipette iji dozie usoro mmeghachi omume ndị a:
| Ngwa | Olu (μL) |
| Ihe nchekwa DNAse I Reaction (10×) | 1 |
| DNasel recombinant (RNase n'efu) | 1 |
| RNA | X |
| ddH enweghị RNase2O | Ruo 10 |
2. The mmeghachi omume ọnọdụ bụ ndị a: 37 ℃, mgbe 15-30 mins, tinye a ikpeazụ ịta nke 2.5 mM EDTA ngwọta na mix ọma, mgbe ahụ 65 ℃ maka 10 mins.Enwere ike iji ndebiri edoziri maka nnwale RT-PCR ma ọ bụ RT-qPCR na-esote, wdg.
Ihe ndetu
1. DNase l na-enwe mmetụta maka denaturation anụ ahụ;Mgbe ị na-agwakọta, jiri nwayọọ tụgharịa tube ule namaa jijiji nke ọma, emela ka ọ maa jijiji nke ukwuu.
2. Ekwesịrị ịchekwa enzyme ahụ n'ime igbe ice ma ọ bụ na mmiri ịsa mmiri mgbe ejiri ya, ma ekwesịrị ịchekwa ya na -20 ℃ ozugbo ejiri ya.
3. Ngwaahịa a bụ maka nyocha naanị.
4. Biko rụọ ọrụ na uwe lab na uwe mkpuchi, maka nchekwa gị.
