Izvo zvinhu zvinosarudza DNA synthesis kunyatsoshanda

Mune yakajairwa DNA, RNA uye isiri-yakasikwa nucleic acids 'synthesis, iyo Deprotection uye Coupling nhanho inoita basa rakakosha.

Danho reDeprotection nderokubvisa boka reDMT parutsigiro rwakasimba kana kuti 5' hydroxyl boka pane yakapfuura nucleoside ine organic acid, uye yakafumura iyo hydroxyl boka reinotevera danho rekubatanidza.Iyo 3% trichloroacetic acid mu dichloromethane kana toluene inonyanya kushandiswa kuita danho rekudzivirira.Iko kusanganiswa kwe trichloroacetic acid uye nguva yekudzivirira (nguva yekubvisa) inotonga kuchena kwezvigadzirwa zvekupedzisira.Kudzika kwakadzika uye kusakwana kwekuvhara nguva inosiya isina kubatwa DMT boka, izvo zvinoderedza goho uye kuwedzera kusvibiswa kusingadiwi.Iyo yakareba yekuvharisa nguva inogona kutungamirira depurine yezvakagadzirwa sequences, kugadzira tsvina isingatarisirwe.

Iyo Coupling nhanho inonzwa kune mvura yemukati yezvinonyungudutsa uye mwando mumhepo.Iko kusanganiswa kwemvura mune synthesis kunofanirwa kunge kuri pasi pe40 ppm, zviri nani isingasviki 25 ppm.Kuchengeta iyo anhydrous synthesis mamiriro, iyo nucleic acids 'synthesis inofanirwa kuitwa munzvimbo yakaderera hunyoro, saka isu tinokurudzira mutengi wedu kushandisaAmidites Yakanyungudika Equipment, iyo inogona kunyungudutsa Phosphoramidite yehupfu kana mafuta mu anhydrous acetonitrile kuti isasangana nemhepo.

Zvinhu zvinotarisa DNA synthesis kunyatsoshanda5
Zvinhu zvinosarudza DNA synthesis kunyatsoshanda4

Sezvo kunyungudika kwephosphoramidites kuri nani pane iyo isiri-mvura mamiriro, uye mamorekuru misungo kuti adsorb iyo inoteedzera mvura mune reagents uye amidite, inofanirwa kugadziriraMolecular Misungo.Tinokurudzira 2 g subsieve ye50-250ml reagent mabhodhoro, 5g ye250-500ml reagent mabhodhoro, 10g yema500-1000ml mabhodhoro reagent, uye 20g yemabhodhoro 1000-2000ml reagent.

Kunyungudika kwephosphoramidites kunofanira kuitwa pasi pemhepo inert, uye kutsiva activator reagents uye acetonitrile kunofanira kupedzwa nenguva.Iyo Capping uye Oxidation reagents inofanirwa kushandiswa nekukurumidza sezvinobvira, iyo yakavhurwa reagents inopa hushoma hupenyu hwesherufu, uye kushoma kuita panguva yekubatanidza.


Nguva yekutumira: Aug-09-2022