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Gịnị bụ Nucleic Acid mmịpụta?
N'okwu kachasị mfe, mmịpụta nucleic acid bụ mwepụ nke RNA na/ma ọ bụ DNA site na nlele na ngafe niile na-adịghị mkpa. Usoro nke mmịpụta na-ekewapụ acid nucleic site na nlele ma na-ewepụta ha n'ụdị eluate gbadoro anya, na-enweghị ihe na-edozi ahụ na ihe mmetọ nke nwere ike imetụta ngwa ọ bụla dị n'okpuru.
Ngwa nke Mwepụta Nucleic Acid
A na-eji acids nucleic dị ọcha mee ihe na plethora nke ngwa dị iche iche, nke sitere na ụlọ ọrụ dị iche iche. Nlekọta ahụike bụ mpaghara ebe a na-ejikarị eme ihe, yana RNA na DNA dị ọcha achọrọ maka ọtụtụ ebumnuche nnwale dị iche iche.
Ngwa nke mmịpụta nucleic acid na ahụike gụnyere:
- Usoro Ọgbọ na-abịa (NGS)
- SNP Genotyping dabere na mmụba
- Genotyping dabere n'usoro
- mgbochi mgbaze enzymes
- Nyochaa site na iji Enzymes na-agbanwe agbanwe (dịka Ligation na cloning)
Enwekwara mpaghara ndị ọzọ gafere ahụike ebe a na-eji mmịpụta nucleic acid, gụnyere mana ọnweghị oke na nnwale nna, ihe nyocha na genomics.
Mkpirikpi akụkọ ihe mere eme nke mmịpụta Nucleic Acid
Mwepụta DNAN'oge dị anya gara aga, ebe otu dọkịta Switzerland aha ya bụ Friedrich Miescher mere mpụ mbụ a mara na 1869. Miescher nọ na-atụ anya idozi ụkpụrụ bụ́ isi nke ndụ site n'ịchọpụta ihe mejupụtara kemịkal nke mkpụrụ ndụ. Mgbe ọ na-adaba na lymphocytes, ọ nwere ike nweta DNA crude site na leukocytes nke a chọtara na pus na bandeeji a tụfuru. O mere nke a site n'itinye acid na alkali na cell ka ọ hapụ cytoplasm cell, wee mepụta usoro iji kewaa DNA na protein ndị ọzọ.
N'ịgbaso nyocha nke Miescher mere n'ala, ọtụtụ ndị sayensị ndị ọzọ aga n'ihu ma mepụta usoro iji kewapụ na sachapụ DNA. Edwin Joseph Cohn, onye ọkà mmụta sayensị protein mepụtara ọtụtụ usoro maka ịdị ọcha protein n'oge WW2. Ọ bụ ya kpatara ikewapụ akụkụ nke serum albumin nke plasma ọbara, nke dị mkpa iji nọgide na-enwe nrụgide osmotic n'ime arịa ọbara. Nke a dị oké mkpa maka idebe ndị agha ndụ.
Na 1953 Francis Crick, ya na Rosalind Franklin na James Watson, kpebisiri ike nhazi nke DNA, na-egosi na ọ bụ eriri abụọ nke ogologo agbụ nke nucleic acid nucleotides mejupụtara ya. Nchọpụta ihe nrịbama a meghere ụzọ maka Meselson na Stahl, bụ ndị nwere ike ịmepụta usoro njupụta gradient centrifugation iji kewapụ DNA na nje bacteria E. Coli ka ha gosipụtara ngbanwe nke DNA na-adịghị agbanwe agbanwe n'oge nnwale 1958 ha.
Usoro nke Nucleic Acid mmịpụta
Kedu usoro 4 nke mmịpụta DNA?
Usoro mmịpụta niile na-agbada n'otu usoro ntọala ahụ.
Mgbochi cell. Oge a, nke a makwaara dị ka cell lysis, gụnyere ịkwatu mgbidi cell na/ma ọ bụ akpụkpọ ahụ cell, iji hapụ mmiri intra-cellular nke nwere acid nucleic nke mmasị.
Mwepụ nke irighiri ihe na-achọghị. Nke a gụnyere lipids akpụkpọ ahụ, protein na acid nucleic ndị ọzọ na-achọghị nke nwere ike igbochi ngwa ala.
Nnọpụ iche. Enwere ụzọ dị iche iche iji kewapụ acid nucleic nke mmasị site na lysate kpochapụrụ nke ị mepụtara, nke na-adaba n'etiti isi ihe abụọ: ngwọta dabeere ma ọ bụ ọnọdụ siri ike (lee ngalaba na-esote).
Ntinye uche. Mgbe e kewapụrụ acids nucleic na ihe ndị ọzọ na-emerụ emerụ na ihe ndị na-edozi ahụ, a na-egosi ha na eluate dị elu.
Ụdị Mịpụta Abụọ Abụọ
Enwere ụdị abụọ nke mmịpụta nucleic acid - ụzọ dabere na ngwọta na ụzọ steeti siri ike. A na-akpọkwa usoro ihe ngwọta dị ka usoro mmịpụta kemịkalụ, ebe ọ na-agụnye iji kemịkalụ na-agbaji cell na ịnweta ihe nucleic. Nke a nwere ike iji ma ogige organic dị ka phenol na chloroform, ma ọ bụ nke na-adịghị emerụ ahụ ya mere ogige inorganic akwadoro dị ka Proteinase K ma ọ bụ silica gel.
Ọmụmaatụ nke ụzọ dị iche iche na-ewepụta kemịkalụ iji gbarie cell gụnyere:
- Osmotic ruptring nke akpụkpọ ahụ
- Enzymatic mgbaze nke cell mgbidi
- Solubilisation nke akpụkpọ ahụ
- Na ncha
- Site na ọgwụgwọ alkali
Usoro steeti siri ike, nke a makwaara dị ka ụzọ igwe, gụnyere iji otu DNA si emekọrịta ihe na mkpụrụ siri ike. Site n'ịhọrọ bead ma ọ bụ molekul nke DNA ga-ejikọta na ya mana onye nyocha agaghị, ọ ga-ekwe omume ikewapụ ha abụọ. Ọmụmatụ nke usoro mmịpụta siri ike nke gụnyere iji silica na bead magnetik.
Akọwara mpụta bead ndọta
Ụzọ ndọpụ bead ndọta
Achọpụtara ikike nke ịwepụta site na iji bead magnetik na patent US nke Trevor Hawkins gbara maka ụlọ ọrụ nyocha nke Whitehead Institute. Ikike ikike ikike a kwetara na ọ ga-ekwe omume iwepụta mkpụrụ ndụ ihe nketa site na ijikọta ha na ihe nkwado siri ike, nke nwere ike ịbụ bead magnet. Ụkpụrụ bụ na ị na-eji bead magnetik arụrụ arụ nke ukwuu nke ihe mkpụrụ ndụ ihe nketa ga-ejikọta na ya, nke enwere ike kewapụ ya na ihe dị elu site n'itinye ike magnetik n'èzí ụgbọ mmiri na-ejide ihe nlele ahụ.
Kedu ihe kpatara eji mwepu bead magnetik?
Nkà na ụzụ mmịpụta bead magnet na-aghọwanye ebe niile, n'ihi ikike ọ nwere maka usoro mmịpụta ngwa ngwa na nke ọma. N'oge na-adịbeghị anya enweela mmepe nke beads magnetik arụ ọrụ nke ukwuu nwere sistemu nchekwa kwesịrị ekwesị, bụ nke mere ka enwere ike ịmepụta mmịpụta acid nucleic yana ọrụ na-arụ ọrụ nke na-arụ ọrụ nke ọma ma na-akwụ ụgwọ. Ọzọkwa, ụzọ ndọta bead adịghị agụnye nzọụkwụ centrifugation nke nwere ike ịkpata ike shiee nke na-agbajisị ogologo DNA. Nke a pụtara na eriri DNA dị ogologo na-adịgide, nke dị mkpa na nyocha nke genomics.
Oge nzipu: Nov-25-2022