Kokubili ukwakheka kwe-DNA kanye ne-RNA kusekelwe isu lokuhlanganisa lesigaba esiqinile kanye namakhemikhali e-phosphoramidite, i-synthesizer ye-DNA ingasetshenziswa ukuze kuhlanganiswe ama-analogue e-RNA noma e-RNA ngaphandle kokuguqulwa okwengeziwe, futhi ama-reagents ku-DNA synthesis angasetshenziswa ngokuqondile ku-RNA kanye nama-nucleic acid okwenziwa. 'i-synthesis.Njengoba kukhonjisiwe kuMfanekiso 1, i-2'-hydroxy ku-RNA phosphoramidite ivikelwe ngeqembu elivikelayo le-silyl, okungukuthi i-t-butyldimethylsilyl (TBDMS), ivimbela ukusabela okusemaceleni eqenjini elibuthakathaka le-2'-hydroxy.Iqembu elikhulu le-TBDMS livimbe ukusabela phakathi kwe-phosphoramidite neqembu le-5'-hydroxy ekusekelweni okuqinile, futhi kudingeka isikhathi eside sokuhlanganisa ukuze kubonakale ukwanelisa ukusebenza kahle kokuhlanganisa.
Ngokufana nokuhlanganiswa kwe-DNA, i-RNA ehlanganisiwe yaqala yahlukaniswa kusukela ekusekelweni okuqinile yi-aminolysis, kwase kuthi iqembu le-TBDMS lahlukaniswa yi-tetrabutylammonium fluoride (TBAF) noma i-trimethylamine trihydrofluoride.I-RNA engahluziwe ingahlanzwa ngokwenziwa kabusha ngotshwala kanye ne-HPLC.
Umfanekiso 1. Isakhiwo samakhemikhali samabhulokhi wokwakha ayisisekelo ku-DNA kanye ne-RNA synthesis.
a) i-dABz phosphoramidite kanye ne-b) i-rABz 2'-OTBDMS phosphoramidite.
Ukwakhiwa komuthi we-siRNA kudinga ama-analogue esakhiwo se-RNA yomdabu ukuze kwandiswe ukuhambelana kwezinto eziphilayo, i-2'-hydroxy ku-RNA ingase ithathelwe indawo yiqembu le-MeO, F kanye ne-MOE, kanti i-nucleic acid ekhiyiwe (LNA) iphinde inikeze ukusebenza kahle ekwelashweni kwe-RNA. (Umfanekiso 2).Lawa ma-phosphoramidite anikeza ukusebenza okufanayo njenge-DNA yohlobo lwe-phosphoramidites ekuhlanganiseni, futhi inqubo yokusebenza nokuhlanza lawa ma-nucleic acid angewona awemvelo afana ne-DNA yomdabu.
Umfanekiso 2. Isakhiwo samakhemikhali samabhulokhi wokwakha emithini ye-siRNA.a) i-dABz 2-MeO phosphoramidite;b) i-dABz 2-F phosphoramidite;c) dABz 2-MOE phosphoramidite kanye d) dABz I-phosphoramidite Ekhiyiwe.
Ama-Phosphoramiditeszibalulekile ekwakhiweni kwezakhi zofuzo, okuhlanganisa ikakhulukazi imindeni ye-DNA kanye ne-RNA kanye nokuphuma kwayo, yonke i-phosphoramidite engenhla esingayinikeza ephaketheni.
Sihlala sithuthukisa ukusebenza kwemikhiqizo yethu, sithuthukisa inqubo yethu yokukhiqiza futhi sithola imininingwane ngendlela efanele.Asikunikezeli ngemikhiqizo yekhwalithi ephezulu kuphela, kodwa futhi sikuhlinzeka ngokuqeqeshwa kanye nenkonzo.Sihlonyiswe ngabasebenzi bokuphatha abangochwepheshe kanye nethimba lezobuchwepheshe elihle kakhulu nabasebenzi bokulungisa ukuze sinikeze amakhasimende ethu isevisi ephelele yangemva kokuthengisa.
Isikhathi sokuthumela: Aug-09-2022