Lako Hoomaemae no ka hoomaemae Oligo

1. Gel Electrophoresis Chromatography Hoomaemae
E hoʻohana i ka polyacrylamide gel electrophoresis chromatography no ka hoʻomaʻemaʻe.ʻO ka denaturing agent ka mea maʻamau 4M formamide a i ʻole 7M urea, ʻo ka neʻe o ka acrylamide ma waena o 5-15%, a ʻo ka hapa o ka methacrylamide ma waena o 2-10%.
Ma hope o ka electrophoresis, pono e hoʻoholo i ke kūlana o ka hui nucleic acid ma lalo o ka irradiation o ke kukui ultraviolet, ʻoki ʻia ka gel e loaʻa ana i ka waikawa nucleic target, wāwahi ʻia ka waikawa nucleic a leached, a laila hoʻopaʻa ʻia ka hopena leaching, desalted, helu ʻia a lyophilized.

2. DMT-On, Hoomaemae HPLC
E koho i ke ʻano DMT-On i ka wā o ka synthesis, ua centrifuged ka hua palau a hoʻopaʻa ʻia i ka lumi wela e wehe i ka amonia nui ma hope o ka aminolysis.
Hoʻokaʻawale ʻia ka hoʻohana ʻana i kahi kolamu C18 me ka acetonitrile a me 10% triethylamine-acetic acid (TEAA) ma ke ʻano he eluent.Ma hope o ka pau ʻana o ka elution, hoʻopaʻa ʻia, a laila wehe ʻia ka hui DMT me ka trifluoroacetic acid.Ma hope o ka hoʻokaʻawale ʻana, hoʻoneʻe ʻia kekahi mau paʻakai a me nā molekole liʻiliʻi ma o kahi paipu i ʻoki ʻia, a hoʻopau ʻia.

Hiki i kēia ala ke loaʻa kahi huahana me ka maʻemaʻe kiʻekiʻe, akā pono e hoʻolohe i ka hiki ʻana o ka depurination.

3. DMT-Off, Hoʻomaʻemaʻe HPLC
E koho i ka DMT-Off i ka wā o ka synthesis, a ua centrifuged ka hua palau a hoʻopaʻa ʻia i ka wela lumi e wehe i ka amonia nui ma hope o ka ammonolysis.
Hoʻokaʻawale ʻia ka hoʻohana ʻana i kahi kolamu C18 me ka acetonitrile a me 10% triethylamine-acetic acid i loko o ka wai e like me ka eluent.Ma hope o ka pau ʻana o ka hoʻokaʻawale ʻana a helu ʻia, ua lyophilized nā aliquot.

Pono kēia ʻano hana i ka hoʻoponopono pono ʻana i nā kūlana kaʻawale, a hiki ke loaʻa pū i nā molekele i hoʻopaʻa ʻia.