Tixanaha genomic ee fayrasyada badankood waa la yaqaan.Baaritaannada aashitada nukliyeerka kuwaas oo ah qaybo gaagaaban oo DNA ah oo loogu talagalay in lagu dhex daro DNA-ga fayraska ama qaybaha RNA.Dareen-celinta silsiladda polymerase (PCR) waa farsamo waxtar badan oo loogu talagalay ogaanshaha fayraska.Hababka ogaanshaha sare ee wax soo saarka ayaa la sameeyay dhawaan.
A. Farsamada hybridization nucleic acid
Isku-dhafka aashitada nukliyeerka, oo inta badan ay ku jiraan qulqulka koonfureed (Koonfurta) iyo Waqooyiga (Waqooyi), waa farsamo cusub oo degdeg ah oo ku saabsan goobta ogaanshaha fayraska.Ujeeddada qiimaynta isku-dhafka ayaa ah in la isticmaalo qaybo gaagaaban oo DNA ah (oo loo yaqaanno “baaritaan”) oo loogu talagalay in lagu dhex milmo DNA-ga fayraska ama qaybaha RNA oo dhammaystiran.Kuleylinta ama daawaynta alkaline-ka, DNA-da laba-geesoon ee bartilmaameedka ah ama RNA ayaa loo kala saarayaa hal xadhig ka dibna waxa lagu dhaqaaqaa taageero adag.Intaa ka dib, baaraha ayaa lagu daraa oo la isku daray DNA ama RNA bartilmaameedka.Sida baaritaanka lagu calaamadiyay isotope ama nuclide aan shucaac ahayn, DNA-da bartilmaameedka ama RNA waxaa lagu ogaan karaa iyada oo loo marayo autoradiography ama nidaamka biotin-avidin.Maadaama inta badan genome-yada fayrasku ay yihiin kuwo iskuxiran oo la isku xigxigay, waxa lagu ogaan karaa iyada oo la adeegsanayo fayraska-taxanayaal gaar ah sida baadhitaannada muunada.Waqtigan xaadirka ah, hababka isku-dhafka waxaa ka mid ah: dhibco dhibco ah, isku-dhafka unugyada unugyada, DNA blotting (DNA) (blot South) iyo RNA blotting (RNA) (Northern blot).
Tignoolajiyada B.PCR
Sanadihii la soo dhaafay, taxanayaal farsamooyin kordhinta nucleic acid ayaa la sameeyay iyadoo lagu saleynayo PCR, si loo tijaabiyo fayrasyada aan dareenka lahayn ama aan la beeran karin.PCR waa hab ka samaysan kara isku xigxiga DNA oo gaar ah iyadoo loo marayo falcelinta in vitro polymerase.Habka PCR waxaa ka mid ah wareegga kulaylka ee saddex tillaabo: denaturation , annealing , iyo kordhinta Heerkulka sare (93 ℃ ~ 95 ℃), DNA-ga laba-geesoodka ah waxaa loo kala saaraa laba qaybood oo DNA ah;ka dibna heerkulka hooseeya (37 ℃ ~ 60 ℃), laba asal oo nucleotide ah oo la isku daray ayaa udub dhexaad u ah qaybaha DNA-da ee dhammaystiran;Halka heerkulka ku habboon ee Taq enzyme (72 ℃), isku dhafka silsiladaha DNA-ga cusub waxay ka bilaabmaan asalka 3'dhamaadka iyadoo la adeegsanayo DNA-ga dhammaystiran sida moodallo iyo hal nucleotides oo ah qalab ahaan.Sidaa darteed wareeg kasta ka dib, hal silsilad DNA ah ayaa lagu kordhin karaa laba silsiladood.Ku celcelinta habkan, silsilad kasta oo DNA ah oo lagu farsameeyey hal wareeg ayaa loo isticmaali karaa qaab-dhismeedka wareegga xiga, tirada silsiladaha DNA-da ayaa la labanlaabmaa wareeg kasta, taas oo macnaheedu yahay soo saarista PCR waxaa lagu kordhiyey xawaaraha 2n.Ka dib 25 ilaa 30 wareegyada, wax soo saarka PCR waxaa lagu gartaa electrophoresis, iyo alaabta DNA ee gaarka ah waxaa lagu arki karaa iftiinka UV (254nm).Faa'iidada gaar ahaaneed, dareenka, iyo ku habboonaanta, PCR waxaa lagu qaatay ogaanshaha bukaan-socodka ee caabuqyada fayraska badan sida HCV, HIV, CMV, iyo HPV.Maadaama PCR uu yahay mid xasaasi ah, waxay ku ogaan kartaa DNA-ga fayraska heerka fg, qalliinka waa in si taxadar leh loo sameeyaa si looga fogaado been-abuurka.Intaa waxaa dheer, natiijada togan ee baaritaanka nucleic acid macnaheedu maaha inuu jiro fayras faafa oo nool oo ku jira muunada.
Isticmaalka ballaaran ee farsamada PCR, farsamooyin iyo habab cusub ayaa la soo saaray iyadoo lagu saleynayo farsamada PCR ujeeddooyin kala duwan oo tijaabo ah.Tusaale ahaan, wakhtiga dhabta ah ee PCR-ga ayaa ogaan kara culayska fayraska;In goobta PCR waxaa loo isticmaalaa in lagu aqoonsado infekshanka fayraska ee unugyada ama unugyada;PCR-ka buulka leh waxa uu kordhin karaa qeexida PCR.Waxaa ka mid ah, PCR-ga tirada dhabta ah ee la soo saaray si degdeg ah.Farsamo badan oo cusub, sida baaritaanka TaqMan hydrolysis, baaritaanka isku-dhafka, iyo baaritaanka molecular beacon probe, ayaa lagu daray waqtiga dhabta ah farsamada PCR ee tirada, taas oo si weyn looga isticmaalo cilmi-baarista caafimaadka.Marka laga soo tago in si sax ah loo aqoonsado culeyska fayraska ee dareeraha jirka bukaanka, habkan ayaa sidoo kale loo isticmaali karaa in lagu ogaado mutant-u-dulqaadka daroogada.Sidaa darteed, PCR-ga tirada dhabta ah waxaa inta badan lagu dabaqaa qiimeynta saameynta daaweynta iyo ilaalinta dulqaadka daroogada.
C. Ogaanshaha sare ee asiidhyada nucleic-ka ee fayraska
Si loo daboolo baahida degdegga ah ee cudurrada faafa ee degdegga ah ee degdegga ah, habab kala duwan oo lagu ogaan karo, sida chips DNA (DNA), ayaa la sameeyay.Chips-yada DNA-da, shaybaarro gaar ah ayaa la farsameeyay oo lagu dhejiyay chips silikoon yar oo cufnaan aad u sarreeya si ay u sameeyaan DNA-da microarray (DNA) kaas oo lagu qasi karo muunad.Calaamadaha isku-dhafka waxaa lagu sawiri karaa mikroskoob confocal ama scanner laser waxaana lagu sii baaraandegi karaa kumbuyuutarka waxaana la heli karaa xog aad u badan oo ku saabsan hiddo-wadaha kala duwan.Waxaa jira laba nooc oo jajab DNA ah."Chip synthesis" waa sida soo socota: oligonucleotides-ka gaarka ah ayaa si toos ah loogu dhejiyay jajabyada.Mid kale waa chip DNA pool.Hidde-sidayaasha cloned ama alaabta PCR ayaa si habsami leh loogu daabacay bogga.Faa'iidada tignoolajiyada chip DNA waa ogaanshaha isku mar ee tiro badan oo taxane DNA ah.Nooca ugu dambeeyay ee jajabka ogaanshaha cudur-sidaha wuxuu aqoonsan karaa in ka badan 1700 fayraska bini'aadamka hal mar.Tiknoolajiyada chip DNA waxay xallisay dhibaatooyinka hababka isku-dhafan ee aashitada nukleic-dhaqameedka waxayna leedahay codsiyo aad u ballaaran oo ku saabsan ogaanshaha fayraska iyo daraasadda cudurrada faafa.
Waqtiga boostada: Dec-23-2020