Lisebelisoa tsa tlhoekiso ea Oligo bakeng sa motsoako oa pele oa Oligo,
mochini oa tšehetso oa oligo synthesis,
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 |
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.
oligo synthesis accerry lisebelisoa bakeng sa dna rna synthesis cycle, e loketseng primer e tloaelehileng, le tse ling tse khutšoanyane tsa ketane, li etsa hore oligo e hloeke haholo.
China moetsi oa lisebelisoa tsa oligo synthesis ka boleng bo botle le tšebeletso e ntle ka ho fetisisa.