DNSase I (Rnase Kyauta)(5u/ul)
Saukewa: HC4007A
DNase I (Deoxyribonuclease I) wani endodeoxyribonuclease ne wanda zai iya narkar da DNA guda ɗaya ko biyu.Yana gane kuma yana raba haɗin phosphodiester don samar da monodeoxynucleotides ko oligodeoxynucleotides guda ɗaya ko biyu tare da ƙungiyoyin phosphate a 5'-terminal da hydroxyl a 3'-terminal.Ayyukan DNAse I ya dogara da Ca2+kuma ana iya kunna su ta hanyar ions na ƙarfe na divalent kamar Mn2+da Zn2+.5mm ku2+yana kare enzyme daga hydrolysis.A gaban Mg2+, Enzym ɗin zai iya ganewa da kayyade kuma ya raba kowane rukunin yanar gizo akan kowane madaidaicin DNA.A gaban Mn2+, za a iya gane igiyoyin DNA guda biyu a lokaci guda kuma a raba su a kusan wuri ɗaya don samar da gutsuttsuran DNA na ƙarshen ƙarewa ko guntuwar DNA na ƙarshe tare da 1-2 nucleotides suna fitowa.
Abubuwan da aka gyara
| Suna | 0.1 KU | 1 KU | 5 KU | 50 KU |
| DNSase I, RNase-free | 20 μl | 200 μL | 1 ml | ml 10 |
| 10×DNase I Buffer | 1 ml | 1 ml | 5 × 1 ml | 5 × 10 ml |
Yanayin ajiya
-25 ℃ ~ -15 ℃ don ajiya;Kai a ƙarƙashin fakitin kankara.
Umarni
1. Shirya maganin amsawa a cikin bututun da ba shi da RNase bisa ga adadin da aka jera a ƙasa:
| Bangaren | Ƙarar |
| RNA | X µg |
| 10 × DNSase I Buffer | 1 μl |
| DNSase I, RNase-kyauta(5U/μL) | 1 U da µg RNA① |
| ddH2O | Har zuwa 10 μL |
Lura: ① Yi ƙididdige ƙarar DNAse I wanda ke buƙatar ƙara dangane da adadin RNA.
2. 37 ℃ na minti 15;
3. Ƙara 0.5M EDTA zuwa ƙaddamarwar ƙarshe na 2.5mM ~ 5mM, da zafi a 65 ℃ na minti 10 don dakatar da amsawa.Za'a iya amfani da samfurin kai tsaye don amsawa ta gaba kamar juyar da rubutugwaji.
Ma'anar Naúrar
An bayyana raka'a ɗaya azaman adadin enzyme wanda zai lalata 1µg na pBR322 gaba ɗayaDNA a cikin minti 10 a 37 ℃.
Kula da inganci
RNase:5U na DNAse I tare da 1.6µg MS2 RNA na awanni 4 a 37℃ ba ya haifar da lalacewa kamarƘaddamar da agarose gel electrophoresis.
Bayanan kula
1. Da fatan za a shirya 0.5MEDTA da kanku.
2. Yi amfani da 1U DNAse I a kowace µg na RNA.Koyaya, idan RNA bai kai 1µg ba, da fatan za a yi amfani da 1U DNAse I.
3. Da fatan za a sanya enzyme akan kankara yayin aiki.
