Ko te whakahiato waikawa karihi ka mahia ma te whakamahi i te waahanga totoka sialic amide triglyceride tikanga, na te mutunga o te 3' o te DNA e kore e nekehia ki runga i te waahanga totoka me te taapiri i nga nucleotides i roto i te ahunga 3' ki te 5' tae noa ki te whakahiato i te wahanga DNA e hiahiatia ana. .He rereke tenei ki te whakahiato DNA ma te whakamahi i te DNA polymerase.
Ko te mutunga o te 3' o te turanga tuatahi ka kore e nekehia i runga i te CPG i te wa o te whakahiato, ko te 5'-OH o te turanga e whai ake nei ka tiakina ki te di-p-tolyl trityl DMT, ko te roopu amino kei runga i te turanga ka tiakina ki te waikawa benzoic me te 3' Ka whakahohehia te -OH me te pūhui amino phosphate.Ko te turanga 1 o te 5 5'-OH o te turanga 1 me te 3'-OH o te turanga e whai ake nei ka hanga he triglyceride phosphite, ka waikurahia ki te iodine ki te triglyceride phosphate, ka tangohia te parenga o te turanga tuarua 5'-OH e te taapiri i nga huringa DM waikawa dichloroacetic na roto i te taapiri o te turanga e whai ake nei, a, i muri i te whakahiato ka tangohia te whakamarumaru i runga i te 5'-OH me te waikawa ngoikore Ka momotuhia te kongakonga mai i te kapia totoka me te haukinia haukinia hydroxide, ka tangohia te whakamarumaru mai i te turanga. me te haukinia haukinia i raro i te whakamahana, ka tangohia te haukinia haukinia, ka whakamarokehia te kongakonga, ka purea te waikawa karihi ma te chromatography wai, PAGE ranei.
Nga taahiraa o te Oligo Synthesis
Werohanga
Ko te5 ka tangohia te roopu whakamarumaru DMT ma te taapiri i te otinga TCA.
Whakahohenga
Whakaranuhia te activator ki te monomer oligonucleotide hei hanga i te oligonucleotide monomer hohe takawaenga.
Te hono
Ka tauhohe te roopu hydroxyl 5-terminal ki te oligonucleotide hohe o te takawaenga i roto i te tauhohenga condensation anti-generative ki te hanga i te herenga phosphite triglyceride iu.
Whakakapi
Ko te taapiri o te kaihoko kapi ka mahia he tauhohenga acetylation capping me te taikaha o nga roopu hydroxyl e kore e uru ki te tauhohenga whakamaarama.
Te hāora
Ko te taapiri o te wairewa iodine kei roto te wai ka tauhohe ki nga here phosphite kore e tau ma te waikura ki te hanga i nga here triglyceride waikawa phosphoric pūmau.
Ko te Cyanoethyl Cleavage
Ko te hua waikawa nucleic i muri i te whakahiatotanga, me te taapiri o te otinga DEA hei tango i te roopu cyanoethyl mai i te herenga triglyceride phosphate.
Werohia me te Tiaki
Ka wehea nga kongakonga waikawa karihi mai i te kaikawe wahanga totoka ma te taapiri i te wairewa haukinia hydroxide katahi ka whakamahana hei tango i nga roopu whakamarumaru mai i nga here hauwai o nga turanga.
Wā tuku: Noema-21-2022