Oligo mea fa'amamāina mo le fa'asologa muamua o Oligo,
masini lagolago oligo synthesis,
laupapa fetaui | 1, 3, 5, 8. |
Filifiliga | fa'amama filtration, suction filtration |
Numera o uafu tui | 5, 6, 7, 8, 9, 10. |
Ituaiga ipu fetaui | C18 ipu, ipu vaieli loloto, ipu fa'apipi'i (feso'ota'i ma le tele o ipu gaosia), ipu microtiter |
Module | tasi axis po o le itu lua |
Malolosi | 220V |
Fa'amaonia | 1 tausaga |
Aganuu | Taliaina |
1. Gel Electrophoresis Chromatography Faʻamamaina
Fa'aaoga le denaturing polyacrylamide gel electrophoresis chromatography mo le fa'amamāina.O le sui denaturing e masani lava o le 4M formamide poʻo le 7M urea, o le faʻaogaina o le acrylamide o loʻo i le va o le 5-15%, ma o le vaega o le methacrylamide e masani lava i le va o le 2-10%.
A maeʻa le electrophoresis, e manaʻomia le faʻamautuina o le tulaga o le nucleic acid band i lalo o le faʻamalamalamaina o le malamalama ultraviolet, o le gel o loʻo i ai le faʻaogaina o le nucleic acid e vavae ese, o le nucleic acid e taʻe ma faʻafefe, ona faʻatumauina lea o le vaifofo, faʻamama, quantified ma lyophilized.
2. DMT-On, HPLC Purification
Filifili le DMT-On mode i le taimi o le faʻapipiʻiina, o le oloa mama sa centrifuged ma faʻaoga i le vevela o le potu e aveese ai le amonia sili atu pe a uma le aminolysis.
O le vavaeeseina na faia i le faʻaaogaina o le koluma C18 faʻatasi ai ma le acetonitrile ma le 10% triethylamine-acetic acid (TEAA) e pei o le eluent.A maeʻa le elution, e faʻapipiʻi, ona aveese lea o le vaega DMT ma le trifluoroacetic acid.A mae'a fa'aleaogaina, e aveese nisi masima ma molelaiti laiti e ala i se paipa tipi, ma i'u ai ina fa'amama.
O lenei metotia e mafai ona maua ai se oloa e sili atu le mama, ae e tatau ona gauai atu i le tupu mai o le depurination.
3. DMT-Off, HPLC Purification
Filifili le DMT-Off i le taimi o le tuʻufaʻatasiga, ma o le oloa faʻatauvaʻa sa centrifuged ma faʻaoga i le vevela o le potu e aveese ai le amonia sili atu pe a uma le ammonolysis.
O le vavaeeseina na faia i le faʻaaogaina o le koluma C18 ma le acetonitrile ma le 10% triethylamine-acetic acid i totonu o le vai e pei o le eluent.A maeʻa ona faʻamaeʻaina ma faʻatusatusaina, o aliquots e lyophilized.
O lenei metotia e manaʻomia ai le faʻaeteete fetuunaiga o tulaga vavae ese, ma e mafai foi ona maua mai mole mole sini mama.
oligo synthesis accerry mea faigaluega mo DNA rna synthesis cycle, lea e fetaui mo le mea masani, ma isi mea fa'apolopolo filifili, faia le oligo maualuga mama.
china gaosi oloa oligo synthesis meafaigaluega ma lelei lelei ma auaunaga sili.