DNase I (Rnase Free) (2u/ul)
Nambala ya mphaka: HC4007B
DNase I ndi endonuclease yomwe imatha kugaya DNA imodzi kapena iwiri.Ikhoza hydrolyze phosphodiester bonds kuti ipange mono-ndi oligodeoxynucleotides yomwe ili ndi gulu la 5'-phosphate ndi gulu la 3'-OH.Mulingo woyenera kwambiri wa pH wa DNase I ndi 7-8.Ntchito ya DNase I imadalira Ca2+ndipo akhoza adamulowetsa ndi divalent zitsulo ayoni monga CO2, Mn2+,zn2+, etc. Pamaso pa Mg2+, DNase Nditha kubala mwachisawawa malo aliwonse a DNA yamitundu iwiri;Ndili pamaso pa Mn2+, DNase Nditha kuphatikizira DNA mozungulira pawiri pamalo amodzi, kupanga malekezero osongoka kapena zomata ndi ma nucleotide 1-2 otulukira.Ikhoza kugwiritsidwa ntchito pokonza zitsanzo zosiyanasiyana za RNA.
Zigawo
| Dzina | 1 KU | 5KU |
| Recombinant DNaseI (RNase-free) | 500 μL | 5 × 500 μL |
| DNase I Reaction Buffer (10×) | 1 ml | 5 × 1 mL |
Zosungirako
Izi ziyenera kusungidwa pa -25 ~ -15 ℃ kwa 2 zaka.Chonde pewani kuzizira kobwerezabwereza.
Malangizo
Amagwiritsidwa ntchito pochotsa DNA kuchokera ku zitsanzo za RNA kuti zingochitika zokha.
1. Chonde gwiritsani ntchito machubu a RNase opanda ma centrifuge ndi malangizo a pipette pokonzekera njira zotsatirazi:
| Zigawo | Kuchuluka (μL) |
| DNase I Reaction Buffer (10×) | 1 |
| Recombinant DNasel (RNase-free) | 1 |
| RNA | X |
| RNase-free ddH2O | Mpaka 10 |
2. Zomwe zimachitika ndi izi: 37 ℃, pambuyo pa 15-30 mins, onjezerani chigawo chomaliza cha 2.5 mM EDTA yankho ndikusakaniza bwino, ndiye 65 ℃ kwa 10 min.Template yokonzedwa itha kugwiritsidwa ntchito pazoyeserera za RT-PCR kapena RT-qPCR, ndi zina.
Zolemba
1. DNase l imakhudzidwa ndi kusintha kwa thupi;Mukasakaniza, pang'onopang'ono sinthani chubu choyesera ndigwedezani bwino, osagwedezeka mwamphamvu.
2. Enzyme iyenera kusungidwa mu bokosi la ayezi kapena pamadzi osambira akagwiritsidwa ntchito, ndipo iyenera kusungidwa pa -20 ℃ mwamsanga mukangogwiritsa ntchito.
3. Mankhwalawa ndi ogwiritsidwa ntchito pofufuza okha.
4. Chonde gwiritsani ntchito malaya a labu ndi magolovesi otayika, kuti mutetezeke.
