ⅠNucleic acidihorudhac
Nucleic acid waxa loo qaybiyaa deoxyribonucleic acid (DNA) iyo ribonucleic acid (RNA), oo ay ka mid yihiin RNA waxa loo qaybiyaa ribosomal RNA(rRNA), messenger RNA (mRNA) iyo wareejinta RNA(tRNA) iyadoo loo eegayo hawlaheeda kala duwan.DNA waxay inta badan ku urursan tahay xudunta, mitochondria iyo chloroform, halka RNA inta badan lagu qaybiyo cytoplasm.Sida aasaaska maadiga ah ee muujinta hidde-sidaha, soo saarista aashitada nucleic waxay ka ciyaartaa door aad muhiim u ah cilmi-baarista bayoolajiga molecular iyo ogaanshaha unugyada unugyada.Feejignaanta iyo nadiifinta soo saarista aashitada nucleic waxay si toos ah u saameyn doontaa PCR-da xiga, isku xigxiga, dhismaha vector, dheefshiidka ensaymka iyo tijaabooyinka kale.
Ⅱ Soo saarista aashitada nukliyeerka iyo habka nadiifinta
① Habka soo saarista phenol/chloroform
Soo saarista phenol/chloroform waa hab qadiimi ah oo loogu talagalay soo saarista DNA-da, taas oo inta badan adeegsata laba dareere organic oo kala duwan si loo daaweeyo muunada, milminta DNA-da nucleic acid ee wejiga biyaha, dufannada wajiga organic, iyo borotiinnada u dhexeeya labada weji.Habkani wuxuu leeyahay faa'iidooyinka qiimaha jaban, nadiif sare iyo saameyn wanaagsan.Khasaarooyinku waa qalliin adag iyo waqti dheer.
② Habka Trizol
Habka Trizol waa habka qadiimiga ah ee soo saarista RNA.Habka Trizol waxaa loo qaybiyaa wajiga aqueous iyo wajiga organic ka dib markii centrifugation leh chloroform, kaas oo RNA waa la kala diri ee wejiga aqueous, wejiga aqueous waxaa loo wareejiyaa tube cusub EP, roobab ayaa la helay ka dib marka lagu daro isopropanol, ka dibna ethanol nadiifinta.Habkani wuxuu ku habboon yahay in RNA laga soo saaro unugyada xayawaanka, unugyada iyo bakteeriyada.
③ Habka sifaynta tiirarka centrifugal
Habka safaynta tiirarka Centrifuge adsorb karaa DNA si gaar ah iyada oo loo marayo qalabka xayeysiis matrix silicon gaar ah, halka RNA iyo borotiinka si habsami leh u gudbi karaan, ka dibna la isticmaalo cusbo sare PH hooseeyo si ay isugu daraan nucleic acid, PH milix hoose elution qiimaha PH si ay u kala soocaan oo loo nadiifiyo nucleic acid.Faa'iidooyinka ayaa ah xoojinta sifaynta sare, xasilloonida sare, looma baahna dareeraha organic, iyo kharash yar.Khasaaraha ayaa ah in ay u baahan tahay in tallaabo-tallaabo lagu saleeyo, tillaabooyin hawlgal oo dheeraad ah.
④ Habka kuul magnetic
Habka tusbaxa magnetic waa in la kala qaybiyo saamiga unugga iyada oo loo marayo lysate, la sii daayo nukleic acid ee muunada, ka dibna molecules nucleic acid ayaa si gaar ah loogu dhejiyaa dusha sare ee birta birta ah, halka wasakhda sida borotiinka iyo sonkorta lagu daayo dareeraha.Iyada oo loo marayo tillaabooyinka kala-goynta unugyada unugyada, birta birta birta ah ee ku xidhan aashitada nucleic, dhaqida acid nucleic, elution nucleic acid, iwm., aashitada nucleic ee saafiga ah ayaa ugu dambeyntii la helay.Faa'iidooyinka ayaa ah hawlgal fudud iyo isticmaalka waqti gaaban, iyada oo aan loo baahnayn tallaabo centrifugation.Waxay leedahay shuruudo farsamo oo hooseeya waxayna xaqiijin kartaa hawlgal toos ah iyo hawlgal ballaaran.Isku darka gaarka ah ee birta birlabeedka iyo aashitada nucleic waxay ka dhigtaa aashitada nukleic ee la soo saaray oo leh feejignaan sare iyo daahirnimo.Khasaaraha ayaa ah in qiimaha suuqa ee hadda jira uu yahay mid qaali ah.
⑤ Hababka kale
Marka lagu daro afarta hab ee kor ku xusan, waxaa jira dildilaacyo karkaraya, habka milixda xoogga leh, habka daawada anionic, habka ultrasonic iyo habka enzymatic, iwm.
Ⅲ Nooca soo saarista aashitada nucleic
Foregene waxay leedahay madal PCR tooska ah ee aduunka hogaaminaysa, labajibbaaran RNA Go'doominta (DNA-keliya + RNA kaliya).Alaabooyinka ugu muhiimsan waxaa ka mid ah xirmooyinka go'doominta DNA/RNA, PCR iyo PCR-ga tooska ah ee dib-u-hawlgalayaasha shaybaadhka unugyada.
① Wadarta soo saarista RNA
Isku geynta shaybaarada soo saarista RNA waxaa ka mid ah dhiiga, unugyada, unugyada xoolaha, dhirta, fayraska, iwm. Nadiifinta sare iyo uruurinta sare ee wadarta guud ee RNA waxaa lagu heli karaa wadarta guud ee soo saarista RNA, taas oo loo isticmaali karo RT-PCR, falanqaynta chip, turjumaada vitro, molecular cloning, Dot Blot iyo tijaabooyin kale.
Foregene la xidhiidhaXirmooyinka Go'doominta RNA
Wadarta Xayawaanka Qalabka Go'doonka RNA--Si dhakhso leh oo hufan uga soo saar wadarta guud ee RNA nadiif-sare iyo tayada sare leh unugyo xayawaanno kala duwan.
Qalabka Go'doominta unugga ee RNA--Wadarta guud ee RNA oo si aad ah loo sifeeyay oo tayo sare leh ayaa laga heli karaa unugyo dhaqan oo kala duwan 11 daqiiqo gudaheed.
Qalabka Go'doominta RNA Wadarta Dhirta--Si degdeg ah uga soo saar wadarta guud ee RNA tayada sare leh shaybaarada dhirta leh polysaccharide hooseeya iyo ka kooban polyphenol.
Qalabka Go'doominta ee RNA--Si degdeg ah uga sooc oo ka nadiifi fayraska RNA muunado ay ka mid yihiin balasmaha, serum, dareeraha jidhka ee ka xorta ah unugyada iyo dhaqamada unugyada sare.
② Soo saarista DNA-da genomic
Saambalada DNA-da genomic waxaa ka mid ah ciidda, saxarada, dhiigga, unugyada, unugyada xoolaha, dhirta, fayraska, iwm -isku xigxiga iyo tijaabooyin kale.
Foregene la xidhiidhaXirmooyinka Go'doominta DNA
Qalabka Go'doominta Unugyada Xoolaha--Sifeynta degdega ah iyo nadiifinta DNA-da genomic ee ilo badan, sida unugyada xayawaanka, unugyada, iwm.
Qalabka DNA ee Midi Kit (1-5ml)--Si dhakhso ah uga nadiifi DNA-ga genomic-ga tayada sare leh ee dhiigga xinjirowga lidka ku ah (1-5ml).
Qalabka Go'doominta ee Kaarka FTA ee Buccal Swab--Si dhakhso ah uga nadiifi DNA-ga genomic-ga tayada sare leh ee shaybaarka kaadhka buccal/FTA
Qalabka Go'doominta DNA-da--Si dhakhso ah u nadiifi oo hel DNA-ga tayada sare leh ee shaybaarada dhirta (oo ay ku jiraan polysaccharides iyo muunadaha dhirta polyphenol)
③ Soo saarista Plasmid
Plasmid waa nooc ka mid ah DNA-da molecule yar oo wareeg ah oo ku jira unugyada, kaas oo ah sidaha caadiga ah ee dib-u-habaynta DNA-da.Habka soo saarista plasmidka waa in laga saaro RNA, laga saaro plasmid ka bakteeriyada genomic DNA, oo laga saaro borotiinka iyo wasakhda kale si loo helo plasmid saafi ah.
Qalabka Guud ee Plasmid Mini--Si dhakhso ah uga nadiifi DNA-da plasmid ee tayada sare leh bakteeriyada la beddelay ee tijaabooyinka bayoolajiga joogtada ah sida isbeddelka iyo dheef-shiidka enzim
④ Noocyada kale ee soo saarista, miRNA saarista, iwm.
Qalabka go'doominta miRNA xayawaanka--Si dhakhso ah oo hufan uga soo saar jajabyada RNA yaryar oo ah 20-200nt miRNA, siRNA, snRNA unugyo iyo unugyo xayawaan oo kala duwan
Ⅳ Shuruudaha soo saarista aashitada nucleic-ka iyo natiijada nadiifintas
① Si loo hubiyo daacadnimada qaabka aasaasiga ah ee nucleic acid.
② Iska yaree faragelinta borotiinka, sonkorta, dufanka iyo macromolecules kale
③ Waa in aanay jirin dareere organic ah ama ion bir ah oo aad u badan oo joojin kara enzyme-ka ku jira muunadaha aashitada nucleic.
④ RNA iyo wasakhowga aashitada nukliyeerka kale waa in la tirtiraa marka la soo saarayo DNA, iyo lidkeeda.
Waqtiga boostada: Nov-24-2022
