Kayan Aikin Tsarkakewa don tsarkakewar Oligo

1. Gel Electrophoresis Chromatography tsarkakewa
Yi amfani da denaturing polyacrylamide gel electrophoresis chromatography don tsarkakewa.Maganin cirewa gabaɗaya shine 4M formamide ko urea 7M, adadin acrylamide yana tsakanin 5-15%, kuma adadin methacrylamide yafi tsakanin 2-10%.
Bayan electrophoresis, ana buƙatar ƙaddara matsayin ƙungiyar nucleic acid a ƙarƙashin hasken ultraviolet, an yanke gel ɗin da ke dauke da acid nucleic da aka yi niyya, an lalatar da nucleic acid kuma an zubar da shi, sa'an nan kuma an tattara maganin leaching, an shafe shi. ƙididdigewa da lyophilized.

2. DMT-On, HPLC tsarkakewa
Zaɓi yanayin DMT-On yayin haɗakarwa, ɗanyen samfurin ya kasance a tsakiya kuma an tattara shi a zafin daki don cire wuce haddi ammonia bayan aminolysis.
An yi rabuwa ta amfani da shafi na C18 tare da acetonitrile da 10% triethylamine-acetic acid (TEAA) a matsayin eluent.Bayan an gama haɓakawa, an tattara shi, sannan an cire ƙungiyar DMT tare da acid trifluoroacetic.Bayan an cire su, ana cire wasu gishiri da ƙananan ƙwayoyin ta hanyar bututun da aka yanke, kuma a ƙarshe a shafe su.

Wannan hanya na iya samun samfurin tare da mafi girman tsarki, amma wajibi ne a kula da abin da ya faru na depurination.

3. DMT-Off, HPLC Tsarkakewa
Zaɓi DMT-Off yayin haɗakarwa, kuma samfurin ɗanyen ya kasance a tsakiya kuma an tattara shi a cikin zafin jiki don cire ammonia da yawa bayan ammonolysis.
An gudanar da rabuwa ta amfani da shafi na C18 tare da acetonitrile da 10% triethylamine-acetic acid a cikin ruwa a matsayin eluent.Bayan an gama rabuwa kuma an ƙididdige su, ana yin lyophilized aliquots.

Wannan hanyar tana buƙatar daidaita yanayin rabuwa a hankali, kuma tana iya samun ingantattun kwayoyin manufa.