I-Poly A Carrier RNA

Inombolo yekati: HC4001A

I-Poly A, i-polyadenylate, iyingxube ye-100 ~ 10000 polyadenylates, eyenziwe i-polymerized nge-polynucleotide phosphorylase in vitro.Ku-vivo, i-poly (a) yengezwa ku-3-terminal ye-mRNA nge-enzyme ukuthuthukisa ukuzinza kwe-mRNA.Ekusetshenzisweni kwe-nucleic acid extraction, ukwengeza i-poly A ku-lysate noma isisombululo esibophezelayo kungathuthukisa isivuno se-DNA ne-RNA.Indlelai-poly A yenza ngcono isivuno se-nucleic acid imi kanje:

1. Ukuxhumana okugcwele ne-adsorption engaphezulu yama-athikili.Ama-athikili amaningi e-polypropylene anogesi omile phezulu, ozokhanga ama-nucleic acid.I-RNA yenkampani yenethiwekhi ingagcwalisa leziimiphumela ye-adsorption futhi yehlise ukulahleka kwethagethi ye-nucleic acid.

2. Yenza ama-trace nuclease angasebenzi: Kunama-nuclease ahlukahlukene kumasampula ezinto eziphilayo kanyeimvelo.I-Poly A ingakwazi ukwenza ama-nuclease alandele umkhondo ezinyathelweni zokuwakhipha noma zokulondoloza ukuzengcono isivuno nokuzinza kwe-target nucleic acids.

3. Ukuncibilika: Esinyathelweni sokuhlanzwa kwe-nucleic acid yemvula elamula noma ebophezelayo, i-poly A ingancipha ne-nucleic acid eqondiwe noma yakhe izinhlayiya ze-polymer ukuzengcono ukululama.

Indlela yokusebenzisa

Thatha inani elifanele lempushana ye-lyophilized, engeza amanzi aphathwe nge-DEPC noma isisombululo sikasawoti we-guanidineyincibilikise ku-0.1-1ug/uL, bese uyipakisha kancane bese uyigcina ku- -20°C.

Izinhlelo zokusebenza

I-1.Virus DNA / RNA extraction: ukwengeza i-1-5ug I-Carrier RNA ku-lysate ingathuthukisa isivuno se-RNA / DNA, iqinise i-nucleic acid ehlosiwe futhi igweme ukuchithwa kwe-nucleic acid ehlanzekile ngesikhathi sokugcina.

2. Ku-micro DNA/RNA isizinda ngendlela ye-membrane yekholomu (<1ug), ukungeza i-RNA yenkampani yenethiwekhi ku-1-5ug kusiza ukuthuthukisa isivuno se-nucleic acid.

3. Ku-alcohol mediated nucleic acid precipitation kanye nesinyathelo sokugxila, ukungezwa kwe-RNA ye-1-2ug yenethiwekhi kuyasiza ukuthuthukisa ukutholakala kwe-RNA yesigaba esifushane.

4. Esixazululweni sokusabela se-PCR se-quantitative probe, ukwengeza i-10-100ng yenethiwekhi ye-RNA kusixazululo sokusabela kuyasiza ukuthuthukisa ukuzwela nokunciphisa i-C._Tinani.