Gabatarwa
Menene Cirin Acid Nucleic?
A cikin mafi sauƙi na sharuddan, haɓakar acid nucleic shine cire RNA da/ko DNA daga samfurin da duk abin da bai dace ba. Tsarin hakar ya keɓe ƙwayoyin nucleic daga samfurin kuma yana samar da su a cikin nau'i na eluate mai daɗaɗɗa, wanda ba shi da kayan maye da gurɓataccen abu wanda zai iya rinjayar kowane aikace-aikacen ƙasa.
Aikace-aikace na Nucleic Acid Extraction
Ana amfani da tsaftataccen acid nucleic a cikin ɗimbin aikace-aikace daban-daban, wanda ya bambanta a cikin masana'antu daban-daban. Kila lafiyar ita ce yankin da aka fi amfani da shi, tare da tsaftataccen RNA da DNA da ake buƙata don ɗimbin dalilai na gwaji daban-daban.
Aikace-aikace na cire nucleic acid a cikin kiwon lafiya sun haɗa da:
- Jeri Mai Gabatarwa (NGS)
- Ƙarfafa tushen SNP Genotyping
- Genotyping na tushen Array
- Ƙuntataccen narkewar Enzyme
- Yana nazarin amfani da Enzymes masu gyara (misali ligation da cloning)
Har ila yau, akwai wasu filayen da suka wuce kiwon lafiya inda ake amfani da hakar acid nucleic, gami da amma ba'a iyakance ga gwajin uba, bincike-bincike da genomics ba.
Takaitaccen Tarihin Hakar Acid Nucleic
Cire DNAAn yi nisa mai nisa, tare da sani na farko da wani likita dan kasar Switzerland mai suna Friedrich Miescher ya yi a shekara ta 1869. Miescher yana fatan warware muhimman ka'idojin rayuwa ta hanyar tantance sinadarai na sel. Bayan ya gaza tare da lymphocytes, ya sami damar samun ɗanyen hazo na DNA daga leukocytes da aka samu a cikin mugunya akan bandages da aka jefar. Ya yi haka ne ta hanyar ƙara acid sannan alkali a cikin tantanin halitta don barin cytoplasm na tantanin halitta, sannan ya samar da wata yarjejeniya ta raba DNA da sauran sunadaran.
Bayan binciken da Miescher ya yi, wasu masana kimiyya da yawa sun ci gaba da haɓaka dabarun keɓewa da tsarkake DNA. Edwin Joseph Cohn, masanin kimiyyar furotin ya haɓaka dabaru da yawa don tsabtace furotin a lokacin WW2. Shi ne ke da alhakin keɓance guntun albumin na jini na jini, wanda ke da mahimmanci don kiyaye matsi na osmotic a cikin tasoshin jini. Wannan yana da mahimmanci don kiyaye sojoji a raye.
A cikin 1953 Francis Crick, tare da Rosalind Franklin da James Watson, sun ƙaddara tsarin DNA, yana nuna cewa an yi shi da nau'i biyu na dogayen sarƙoƙi na nucleotides na nucleic acid. Wannan ci gaban binciken ya ba da hanya ga Meselson da Stahl, waɗanda suka sami damar haɓaka ƙa'idar centrifugation mai yawa don ware DNA daga ƙwayoyin E. Coli yayin da suka nuna kwafin DNA na ɗan ra'ayin mazan jiya yayin gwajin 1958.
Dabarun Hako Acid Nucleic
Menene matakai 4 na hakar DNA?
Duk hanyoyin cirewa suna gangara zuwa matakai na asali iri ɗaya.
Rushewar salula. Wannan mataki, wanda kuma aka sani da sel lysis, ya haɗa da rushe bangon tantanin halitta da/ko membrane na tantanin halitta, don saki ruwan ciki na cikin salula wanda ke dauke da acid nucleic na sha'awa.
Cire tarkacen da ba'a so. Wannan ya haɗa da lipids na membrane, sunadarai da sauran acid nucleic maras so waɗanda zasu iya tsoma baki tare da aikace-aikacen ƙasa.
Kaɗaici. Akwai hanyoyi daban-daban da yawa don keɓance acid ɗin nucleic na sha'awa daga sharertaccen lysate da kuka ƙirƙira, wanda ya faɗi tsakanin manyan nau'ikan guda biyu: tushen bayani ko ingantaccen yanayi (duba sashe na gaba).
Hankali. Bayan an ware acid ɗin nucleic daga duk sauran gurɓatattun abubuwa da abubuwan diluents, an gabatar da su a cikin eluate mai mahimmanci.
Nau'u Biyu Na Hakowa
Akwai nau'o'i biyu na hakar acid nucleic - hanyoyin tushen bayani da kuma tsauraran hanyoyin jihar. Hanyar tushen bayani kuma ana kiranta da hanyar hako sinadarai, kamar yadda ya ƙunshi amfani da sinadarai don wargaza tantanin halitta da samun damar abun cikin tsakiya. Wannan na iya zama ta amfani da ko dai kwayoyin halitta irin su phenol da chloroform, ko mafi ƙarancin cutarwa don haka ƙarin shawarwarin mahaɗan inorganic kamar Proteinase K ko gel silica.
Misalan hanyoyi daban-daban na hakar sinadarai don rushe tantanin halitta sun haɗa da:
- osmotic rupturing na membrane
- Enzymatic narkewa na cell bango
- Solubilisation na membrane
- Tare da wanki
- Tare da maganin alkali
Ƙaƙƙarfan fasaha na ƙasa, wanda kuma aka sani da hanyoyin inji, ya haɗa da yin amfani da yadda DNA ke mu'amala da ƙaƙƙarfan ma'auni. Ta zaɓin dutsen dutse ko kwayoyin halitta wanda DNA zai ɗaure a kai amma mai binciken ba zai yi ba, yana yiwuwa a raba su biyun. Misalai na fasaha mai ƙarfi-tsara gami da amfani da silica da beads na maganadisu.
An Bayyana Haɗin Ƙaƙwalwar Ƙwallon Ƙwallon Ƙwallon Ƙwallon Ƙwallon Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwaƙwalwa na Ƙaƙwalwa na Ƙaƙwalwa na Ƙaƙwalwa na Ƙaƙwalwa na Magnetic Bead ) na Ƙuƙwalwa na Magnetic Bead ya yi ya bayyana
Hanyar Haɓakar Ƙwallon Ƙwallon Ƙwallon Ƙwallon Ƙwallon Ƙwallon Ƙwaƙwalwa
An fara gane yuwuwar hakowa ta amfani da beads na maganadisu a cikin takardar izinin Amurka ta Trevor Hawkins, don cibiyar bincike ta Whitehead. Wannan haƙƙin mallaka ya yarda cewa yana yiwuwa a fitar da kwayoyin halitta ta hanyar ɗaure su zuwa wani ƙaƙƙarfan mai ɗaukar hoto, wanda zai iya zama dutsen maganadisu. Ka'idar ita ce, kuna amfani da ƙwanƙolin maganadisu mai aiki sosai wanda kwayoyin halitta za su ɗaure a kai, wanda za'a iya raba shi da maɗaukakiyar ta hanyar amfani da ƙarfin maganadisu zuwa wajen jirgin da ke riƙe da samfurin.
Me yasa Ake Amfani da Haɗin Ƙaƙwalwar Ƙwaƙwalwa?
Fasahar cire katako na Magnetic tana ƙara yaɗuwa, saboda yuwuwar da take da shi na hanyoyin fitar da sauri da inganci. A cikin 'yan lokutan nan an sami ci gaba na kayan aikin maganadisu masu aiki sosai tare da tsarin buffer masu dacewa, waɗanda suka yi yuwuwar sarrafa sarrafa acid nucleic da aikin aiki wanda ke da haske na albarkatu da tsada. Har ila yau, hanyoyin da ake hako dutsen maganadisu ba su haɗa da matakan ɗaiɗaikun ɗaiɗaikun ɗaiɗaikun ɗaiɗaikun ɗaiɗaikun ɗaiɗaikun ɗaiɗaikun ɗaiɗaikun ɗaiɗaikun ɗaiɗaikun ɗaiɗaikun ɗaiɗaikun ɗaiɗaikun ɗaiɗaiɗai waɗanda ke wargaza dogon guntun DNA ba. Wannan yana nufin cewa dogayen igiyoyin DNA sun kasance cikakke, wanda ke da mahimmanci a gwajin kwayoyin halitta.
Lokacin aikawa: Nuwamba-25-2022