Lisebelisoa tsa ho hloekisa bakeng sa tlhoekiso ea oligo
Tlhaloso
| Liboto tse lumellanang | 1, 3, 5, 8. |
| Tlhophiso | ho tlhotla, ho monya |
| Palo ea likou tsa ente | 5, 6, 7, 8, 9, 10. |
| Mefuta ea lipoleiti tse lumellanang | Letlapa la C18, poleiti e tebileng ea seliba, poleiti ea maiketsetso (e lumellana le lipoleiti tse ngata tsa maiketsetso), poleiti ea microtiter |
| Mojule | mokoloko o le mong kapa mokoloko o kopane |
| Palo ea li-volts | 220V |
| Tiisetso | 1 selemo |
| Tloaelo | E amohetsoe |
Mekhoa e fapaneng ea ho hloekisa
1. Gel Electrophoresis Chromatography Hloekiso
Sebelisa denaturing polyacrylamide gel electrophoresis chromatography bakeng sa tlhoekiso.The denaturing agent ka kakaretso ke 4M formamide kapa 7M urea, concentration ea acrylamide e lipakeng tsa 5-15%, 'me karolo ea methacrylamide haholo-holo e lipakeng tsa 2-10%.
Ka mor'a electrophoresis, boemo ba sehlopha sa nucleic acid bo hloka ho khethoa tlas'a mahlaseli a mahlaseli a ultraviolet, gel e nang le sepheo sa "nucleic acid" e khaoloa, "nucleic acid" e pshatleha 'me e tšeloa, ebe tharollo ea leaching e tsepamisitsoe, e tlosoa letsoai. quantified le lyophilized.
2. DMT-On, HPLC Hloekiso
Khetha mokhoa oa DMT-On nakong ea ho kopanya, sehlahisoa se tala se ne se le centrifuged 'me se tsepamisitsoe mocheso oa kamore ho tlosa ammonia e feteletseng ka mor'a aminolysis.
Karohano e ne e etsoa ho sebelisoa mohala oa C18 o nang le acetonitrile le 10% triethylamine-acetic acid (TEAA) e le tlhaloso.Ka mor'a hore tlhaloso e phethoe, e tsepamisitsoe, ebe sehlopha sa DMT se tlosoa ka trifluoroacetic acid.Ka mor'a ho se nke lehlakore, matsoai a mang le limolek'hule tse nyenyane li ntšoa ka tube e khaotsoeng, 'me qetellong e tlosoa letsoai.
Mokhoa ona o ka fumana sehlahisoa se nang le bohloeki bo phahameng, empa hoa hlokahala ho ela hloko ketsahalo ea depurination.
3. DMT-Off, HPLC Hloekiso
Khetha DMT-Off nakong ea ho kopanya, 'me sehlahisoa se tala se ne se le centrifuged' me se tsepamisitsoe mocheso oa kamore ho tlosa ammonia e feteletseng ka mor'a ammonolysis.
Karohano e ne e etsoa ho sebelisoa kholumo ea C18 e nang le acetonitrile le 10% ea triethylamine-acetic acid ka metsing e le mokhoa o hlakileng.Ka mor'a hore karohano e phethoe le ho lekanngoa, li-aliquots li na le lyophilized.
Mokhoa ona o hloka phetoho e hlokolosi ea maemo a karohano, hape o ka fumana limolek'hule tse batlang li hloekile.
