Hauv cov DNA, RNA thiab non-natural nucleic acids 'synthesis, Deprotection thiab Coupling kauj ruam plays lub luag haujlwm tseem ceeb.
Cov kauj ruam Deprotection yog tshem tawm DMT pawg ntawm cov khoom txhawb nqa lossis 5 'hydroxyl pawg ntawm cov nucleoside yav dhau los nrog cov organic acid, thiab nthuav tawm cov pab pawg hydroxyl rau cov kauj ruam nram qab no.3% trichloroacetic acid nyob rau hauv dichloromethane los yog toluene feem ntau yog siv los ua cov kauj ruam tiv thaiv.Lub concentration ntawm trichloroacetic acid thiab lub sij hawm deprotection (deblocking lub sij hawm) dominate lub purity ntawm cov khoom kawg.Tsawg concentration thiab tsis txaus deblocking lub sij hawm tawm unreacted DMT pab pawg neeg, uas txo cov yield thiab nce lub undesired impurities.Lub sij hawm deblocking ntev yuav ua rau cov depurine ntawm synthesized sequences, tsim cov impurities npaj txhij.
Lub Coupling kauj ruam yog rhiab heev rau cov dej ntsiab lus ntawm cov kuab tshuaj thiab cov dej noo hauv huab cua.Qhov concentration ntawm cov dej nyob rau hauv lub synthesis yuav tsum tsawg tshaj li 40 ppm, zoo dua tsawg tshaj li 25 ppm.Txhawm rau ua kom cov txheej txheem anhydrous synthesis, cov nucleic acids 'synthesis yuav tsum tau ua nyob rau hauv qhov chaw tsis muaj av noo, yog li peb xav kom peb cov neeg siv khoom siv cov khoom siv.Amidites Dissolved Equipment, uas muaj peev xwm yaj cov hmoov los yog oily Phosphoramidite nyob rau hauv anhydrous acetonitrile kom tsis txhob sib cuag nrog huab cua.
Txij li thaum yaj ntawm phosphoramidites nws zoo dua nyob rau hauv qhov teeb meem tsis yog dej, thiab molecular ntxiab rau adsorb cov dej nyob rau hauv cov reagents thiab amidite, nws yuav tsum tau npaj lubMolecular Traps.Peb pom zoo 2 g subsieve rau 50-250ml reagent fwj, 5g rau 250-500ml reagent fwj, 10g rau 500-1000ml reagent fwj, thiab 20g rau 1000-2000ml reagent fwj.
Cov yaj ntawm phosphoramidites yuav tsum tau nqa tawm nyob rau hauv lub inert cua, thiab hloov cov activator reagents thiab acetonitrile yuav tsum tiav nyob rau hauv lub sij hawm.Lub Capping thiab Oxidation reagents yuav tsum tau siv sai li sai tau, cov reagents qhib muab lub neej txee tsawg dua, thiab kev ua haujlwm tsawg dua thaum lub sijhawm ua ke.
Post lub sij hawm: Aug-09-2022