Phapang pakeng tsa DNA le RNA synthesis

Ka bobeli DNA le RNA synthesis e ipapisitse le leano le tiileng la phase le chemistry ea phosphoramidite, synthesizer ea DNA e ka sebelisoa ho kopanya li-analogue tsa RNA kapa RNA ntle le phetoho e ngoe hape, 'me li-reagents tse ho DNA synthesis li ka sebelisoa ka kotloloho ho RNA le li-nucleic acid tsa maiketsetso. 'moho.Joalokaha ho bontšitsoe setšoantšong sa 1, 2'-hydroxy ho RNA phosphoramidite e sirelelitsoe ka sehlopha se sireletsang silyl, ke hore, t-butyldimethylsilyl (TBDMS), ho thibela liketso tse ka thōko ho sehlopha se fokolang sa 2'-hydroxy.Sehlopha se seholo sa TBDMS se sitisitse karabelo lipakeng tsa phosphoramidite le sehlopha sa 5'-hydroxy mabapi le ts'ehetso e tiileng, mme ho hlokahala nako e telele ea ho kopanya ho fihlela katleho ea khokahanyo.

Joalo ka ha DNA synthesis, RNA e kopaneng e ile ea qala ho ntšoa ho tsoa ts'ehetso e tiileng ke aminolysis, 'me sehlopha sa TBDMS se ile sa khaoloa ke tetrabutylammonium fluoride (TBAF) kapa trimethylamine trihydrofluoride.RNA e sa jeseng litheohelang e ka hloekisoa ka ho nchafatsoa joala le HPLC.

DNA le RNA synthesis1

Setšoantšo sa 1. Sebopeho sa lik'hemik'hale sa li-block tsa motheo tsa mohaho ho DNA le RNA synthesis.

a) dABz phosphoramidite le b) rABz 2'-OTBDMS phosphoramidite.
Nts'etsopele ea moriana oa siRNA e hloka sebopeho sa li-analogues tsa RNA ea matsoalloa ho eketsa biocompatibility, 2'-hydroxy ho RNA e ka nkeloa sebaka ke sehlopha sa MeO, F le MOE, 'me nucleic acid e notletsoeng (LNA) le eona e fana ka ts'ebetso e ntle kalafong ea RNA. (Setšoantšo sa 2).Li-phosphoramidite tsena li fana ka mesebetsi e tšoanang le ea DNA ea mofuta oa phosphoramidite ka har'a motsoako, 'me ts'ebetso ea ho sebetsa le ho hloekisa li-nucleic acid tsena tseo e seng tsa tlhaho li tšoana le tsa DNA ea tlhaho.

DNA le RNA synthesis2

Setšoantšo sa 2. Sebopeho sa lik'hemik'hale sa li-block tsa ho haha ​​​​litlhare tsa siRNA.a) dABz 2-MeO phosphoramidite;b) dABz 2-F phosphoramidite;c) dABz 2-MOE phosphoramidite le d) dABz E notletsoe phosphoramidite.

Phosphoramiditesli bohlokoa bakeng sa ho qaptjoa ha liphatsa tsa lefutso, ho kenyelletsa haholo-holo malapa a DNA le RNA le lihlahisoa tsa tsona, phosphoramidite eohle e ka holimo eo re ka e fanang ka eona ka har'a sephutheloana.

Re lula re ntlafatsa ts'ebetso ea lihlahisoa tsa rona, re ntlafatsa ts'ebetso ea rona ea tlhahiso le ho fumana lintlha tse nepahetseng.Ha re u fe lihlahisoa tsa boleng bo holimo feela, empa hape re u fa koetliso le tšebeletso.Re na le basebetsi ba botsamaisi ba litsebi le sehlopha se hloahloa sa tekheniki le basebetsi ba tlhokomelo ho fa bareki ba rona ts'ebeletso e phethahetseng ea kamora ho rekisa.


Nako ea poso: Aug-09-2022