Izinsiza Zokuhlanzwa zokuhlanzwa kwe-Oligo
Ukucaciswa
| Amabhodi ahambisanayo | 1, 3, 5, 8. |
| Ukuhlunga | ukuhlungwa kwe-blow, ukuhlunga kokudonsa |
| Inombolo yezimbobo zokujova | 5, 6, 7, 8, 9, 10. |
| Izinhlobo zamapuleti ahambisanayo | Ipuleti le-C18, ipuleti elijulile, ipuleti lokwenziwa (lihambisana namapuleti amaningi okwenziwa), ipuleti le-microtiter |
| Imojuli | i-eksisi eyodwa noma i-eksisi ekabili |
| Amandla kagesi | 220V |
| Iwaranti | 1 unyaka |
| Ngokwezifiso | Kwamukelwe |
Izindlela ezahlukene zokuhlanza
1. Gel Electrophoresis Chromatography Ukuhlanzwa
Sebenzisa i-denaturing polyacrylamide gel electrophoresis chromatography ukuze uhlanzeke.I-ejenti ye-denaturing ngokuvamile i-4M formamide noma i-7M urea, ukugxila kwe-acrylamide kuphakathi kuka-5-15%, futhi ingxenye ye-methacrylamide iphakathi kuka-2-10%.
Ngemuva kwe-electrophoresis, indawo yebhande le-nucleic acid idinga ukunqunywa ngaphansi kwemisebe yokukhanya kwe-ultraviolet, ijeli equkethe i-nucleic acid ehlosiwe iyanqanyulwa, i-nucleic acid iphihlizwe futhi ifakwe, bese isixazululo se-leaching sigxiliwe, sikhishwe usawoti, quantified futhi lyophilized.
2. I-DMT-On, i-HPLC Purification
Khetha imodi ye-DMT-On ngesikhathi sokuhlanganiswa, umkhiqizo ongahluziwe wawufakwe phakathi futhi ugxiliswe ekamelweni lokushisa ukuze kukhishwe i-ammonia eningi ngemva kwe-aminolysis.
Ukuhlukaniswa kwenziwa kusetshenziswa ikholomu ye-C18 ene-acetonitrile kanye ne-10% ye-triethylamine-acetic acid (TEAA) njenge-elient.Ngemva kokuqedwa kwe-lution, kuyagxiliswa, bese iqembu le-DMT lisuswa nge-trifluoroacetic acid.Ngemuva kwe-neutralization, amanye ama-salts nama-molecule amancane akhishwa ngeshubhu elinqunyiwe, futhi ekugcineni akhishwe usawoti.
Le ndlela ingathola umkhiqizo ngokuhlanzeka okuphezulu, kodwa kuyadingeka ukuba unake ukwenzeka kwe-depurination.
3. I-DMT-Off, i-HPLC Purification
Khetha i-DMT-Off ngesikhathi sokuhlanganiswa, futhi umkhiqizo ongahluziwe wawugxiliswe phakathi futhi ugxiliswe ekamelweni lokushisa ukuze kukhishwe i-ammonia eyengeziwe ngemva kwe-ammonolysis.
Ukwehlukaniswa kwenziwa kusetshenziswa ikholomu ye-C18 ene-acetonitrile kanye ne-10% ye-triethylamine-acetic acid emanzini njengokulula.Ngemuva kokuthi ukuhlukaniswa kuqediwe futhi kunqunywe, ama-aliquots a-lyophilized.
Le ndlela idinga ukulungiswa ngokucophelela kwezimo zokuhlukaniswa, futhi ingathola nama-molecule ahlosiwe amsulwa.
