Dhowr hindise oo kacaan ah oo ku jira taariikhda tignoolajiyada ogaanshaha ee maskaxdayda ayaa ah tignoolajiyada difaaca jirka ee ku salaysan mabda'a antigen-antibody xidhid gaar ah, tignoolajiyada PCR iyo tignoolajiyada isku xigxiga.Maanta waxaan ka hadli doonaa farsamada PCR.Marka loo eego horumarka tignoolajiyada PCR, dadku waxay caado ahaan u qaybiyaan tignoolajiyada PCR saddex jiil: tignoolajiyada PCR caadiga ah, tignoolajiyada PCR-ga ee fluorescent-ka iyo farsamada PCR dhijitaalka ah.
Cfarsamada PCR ommon
KARY MULLIS (1944.12.28-2019.8.7)
Kary Mullis waxa uu ikhtiraacay falcelinta silsiladda polymerase (polymerase chain reaction, PCR) 1983. Waxaa la sheegay in markii uu gaadhiga waday saaxiibtiis, uu si lama filaan ah u soo booday waxyi iyo fikir ku saabsan mabda'a PCR ( faa'iidooyinka wadista baabuurta).Kary Mullis waxa lagu abaalmariyay bilada Nobel Prize ee Kimistariga 1993. The New York Times ayaa faallo ka bixisay: “Asal ahaan aad u weyn oo muhiim ah, ku dhawaad u qaybinta bayoolajiga pre-PCR iyo post-PCR.
Mabda'a PCR: Under catalysis ee polymerase DNA, hooyada strand DNA loo isticmaalo sida template ah, iyo primer gaar ah loo isticmaalo sida barta bilowga ah ee kordhinta, iyo strand inanta DNA kaaba in hooyada strand template DNA waxaa lagu koobiyeeyay in vitro iyada oo loo marayo denaturation, annealing, fidinta iyo tallaabooyin kale.Waa tignoolajiyada kordhinta isku-darka DNA-ga ee vitro, taas oo si degdeg ah oo gaar ah u kordhin karta DNA kasta oo la beegsanayo ee ku jirta vitro.
Faa'iidooyinka PCR caadiga ah
1.Habka caadiga ah, heerarka caalamiga ah iyo kuwa gudaha oo dhamaystiran
2.Qiimaha hoose ee reagents qalab
3.Alaabooyinka PCR waxaa lagu soo ceshan karaa tijaabooyinka kale ee bayoolaji ee unugyada
Mashiinka Foregene PCR ee lagu taliyay: https://www.foreivd.com/foreamp-sn-695-series-thermal-cycler-96-wells-pcr-machine-product/
Alaabooyinka la xiriira: https://www.foreivd.com/pcr-herotm-with-dye-product/
Khasaaraha PCR caadiga ah
1.fududahay in la wasakheeyo
2.qalliin adag
3.kaliya falanqaynta tayada
4.Dareen dhexdhexaad ah
5.Waxaa jira cod-weyneyn aan gaar ahayn, marka kooxda aan gaarka ahayn ay la mid tahay xajmiga bartilmaameedka, lama kala saari karo
CPCR ku salaysan apillary electrophoresis
Iyadoo laga jawaabayo cilladaha PCR-ga caadiga ah, qaar ka mid ah wax-soo-saarayaasha ayaa soo bandhigay qalab ku salaysan mabda'a electrophoresis capillary.Talaabada electrophoresis ka dib kordhinta PCR ayaa lagu dhammeeyaa xididka.Dareenku wuu sarreeyaa, faraqa u dhexeeya dhowr saldhigna waa la kala saari karaa, kordhinta waxaa xisaabin kara MAERKER.content alaabta.Khasaaraha ayaa ah in badeecada PCR ay wali u baahan tahay in la furo oo la geliyo qalabka, waxaana wali jirta khatar weyn oo faddarayn ah.
CapillaryElectrophoresis
2. Farsamada tirooyinka PCR ee wakhtiga-dhabta ah (Quantitative Real-time PCR, qPCR)Fluorescent quantitative PCR, oo sidoo kale loo yaqaan Real-Time PCR, waa tikniyoolajiyad tiro cusub oo nucleic acid ah oo ay soo saartay PE (Perkin Elmer) 1995. Taariikhda horumarka ee PCR tirooyinka fluorescent waa taariikhda halganka naf-kicinta ee Rafaa sida ABI, Roche, iyo Bio-Rad.Haddii aad xiisaynayso, waad hubin kartaa.Farsamadani hadda waa farsamada PCR-ta ugu qaan-gaadhka badan uguna isticmaalka badan.
Mashiinka qPCR ee lagu taliyay:https://www.foreivd.com/mini-real-time-pcr-system-forequant-sf2sf4-product/
Habka dheeha fluorescent (SYBR Cagaaran I):SYBR Cagaaran I waa dheeha DNA-da ee inta badan loo isticmaalo PCR-ga tirada badan, kaas oo si aan gaar ahayn ugu xidha DNA-ga laba-geesoodka ah.Gobolka xorta ah, SYBR Green waxay soo saartaa iftiin daciif ah, laakiin marka lagu xidho DNA-ga laba-geesoodka ah, iftiinkeedu wuxuu kordhiyaa 1000-laab.Sidaa darteed, wadarta calaamada fluorescent ee ay soo saarto fal-celintu waxay la mid tahay cadadka DNA-da laba-geesoodka ah ee jooga waxayna kordhin doontaa korodhka badeecada la xoojiyay.Maadaama dheehu si gaar ah ugu xidho DNA-da laba-geesoodka ah, natiijooyin been ah ayaa laga yaabaa in la soo saaro.
Alaabooyinka la xiriira: https://www.foreivd.com/real-time-pcr-easytm-sybr-green-i-kit-product/
Habka baaritaanka fluorescent (teknoolajiyada Taqman): Inta lagu jiroKordhinta PCR, baaritaan gaar ah oo fluorescent ah ayaa lagu daraa isla mar ahaantaana lamaanaha aasaasiga ah.Baadhitaanku waa oligonucleotide toosan, oo leh koox wariyeyaal fluorescent ah iyo koox dab-demiye ah oo ay u kala horreeyaan ku calaamadsan labada daraf.Marka baaritaanka uu yahay mid sax ah, calaamada fluorescent ee ay sii daayaan kooxda weriyaha waxaa nuugaya kooxda quencher, iyo ogaanshaha Ma jiro calaamad fluorescent ah;inta lagu guda jiro kor u qaadida PCR (marxaladda fidinta), 5'-3' Dhaqdhaqaaqa Dicer ee Taq enzyme ayaa dheefshiidi doonta oo hoos u dhigi doonta baaritaanka, si markaa kooxda fluorescent weriyaha iyo kooxda fluorescent quencher la kala saaro, si nidaamka kormeerka fluorescent signalka waa la heli karaa, taas oo ah, mar kasta oo silsilad DNA ah la kordhiyo, calaamad muujinaysa molecule oo dhammaystiran oo la sameeyay molecule fluorescent. s iyo samaynta alaabta PCR.Habka baaritaanka Taqman waa habka ogaanshaha inta badan loo isticmaalo ogaanshaha bukaan-socodka.
Alaabooyinka la xiriira: https://www.foreivd.com/quickeasy%e1%b5%80%e1%b4%b9-real-time-pcr-kit-taqman-product/
Faa'iidooyinka qPCR
1.Habka waa qaan oo qalabkii taageerada iyo reagents waa dhamaystiran
2.Qiimaha dhexdhexaadka ah ee reagents
3.fududahay in la isticmaalo
4.Dareenka ogaanshaha sare iyo gaar ahaaneed
Khasaaraha qPCR
Isbeddelka hidde-sidaha bartilmaameedku wuxuu horseedaa ogaanshaha la seegay.
Natiijada ogaanshaha qaab-dhismeedka hoose lama go'aamin karo.
Waxaa jira qalad weyn marka la isticmaalayo qalooca caadiga ah ee ogaanshaha tirada.
3. Farsamada PCR Digital (digital PCR, dPCR).
PCR-ga dhijitaalka ah waa farsamo lagu qiyaaso molecules acid nucleic.Marka la barbar dhigo qPCR, PCR dhijitaalka ah waxay si toos ah u akhrin kartaa tirada DNA/RNA molecules, taas oo ah qiyaasta buuxda ee unugyada nucleic acid ee muunadda bilowga ah.1999kii, Bert Vogelstein iyo Kenneth W. Kin-zler waxay si rasmi ah u soo jeediyeen fikradda dPCR.
Sannadkii 2006, Fluidigm wuxuu ahaa kii ugu horreeyay ee soo saara qalab dPCR-ku-saleysan ganacsi.Sannadkii 2009, Tiknoolajiyada Nolosha waxay bilawday nidaamyada OpenArray iyo QuantStudio 12K Flex dPCR.2013, Tiknoolajiyada Nolosha ayaa bilaabay nidaamka QuantStudio 3DdPCR, kaas oo adeegsada tignoolajiyada cufnaanta sare ee nanoscale microfluidic chip si ay si siman u qaybiso muunadaha 20,000 unug shaqsiyeed.falcelinta si fiican.
Sannadkii 2011, Bio-Rad waxay soo saartay qalabka QX100 dPCR ee ku salaysan dhibicda, kaas oo adeegsada tignoolajiyada biyaha-saliidka si ay si siman u qaybiso muunadda ilaa 20,000 dhibic-biyo-saliid ah, waxayna isticmaashaa falanqeeye dhibicda si loo falanqeeyo dhibcaha.Sannadkii 2012, RainDance waxay soo saartay qalabka RainDrop dPCR, oo uu kaxeeyo gaaska cadaadiska sarreeya, si loo qaybiyo nidaam kasta oo falcelin caadi ah oo falcelin ah oo ka kooban 1 milyan ilaa 10 milyan oo dhibic-dhibcood ah oo picoliter ah.
Ilaa hadda, PCR dhijitaalka ah wuxuu sameeyay laba qaybood oo waaweyn, nooca chip iyo nooca dhibicda.Si kastaba ha noqotee nooca PCR dhijitaalka ah, mabaadi'da asaasiga ah ayaa xaddidaya milanka, PCR dhammaadka iyo qaybinta Poisson.Nidaamka falcelinta PCR-ga caadiga ah ee ka kooban qaababka nucleic acid waxaa si siman loo qaybiyaa tobanaan kun oo falcelin PCR ah, kuwaas oo loo qaybiyo chips ama microdroplets, si fal-celin kasta uu ka kooban yahay inta ugu badan ee suurtogalka ah molecule template ah, iyo falcelinta PCR hal-molecule ayaa la sameeyaa.Marka la akhriyo fluorescence Joogitaanka ama maqnaanshaha calaamada waa la tiriyaa, waxaana qiyaasida buuxda la sameeyaa ka dib marka la qiyaaso qaybinta Poisson ee tirakoobka.
Kuwa soo socda waa sifooyinka dhowr aaladood oo PCR dhijitaalka ah oo aan isticmaalay:
1. Bio-Rad QX200 dhibicda dhijitaalka ah PCR Bio-RadQX200 waa madal PCR dhijitaalka ah oo aad u heersaraysa, habka ogaanshaha aasaasiga ah: 20,000 muunado waxa soo saara koronto dhaliyaha dhibicda biyaha ku jira ee saliidda ah ayaa lagu kordhiyey mishiinka PCR caadiga ah, ugu dambayntiina calaamadda fluorescence ee dhibic kasta oo yar waxa akhriya akhristaha dhibic-yar.Hawlgalku aad ayuu u dhib badan yahay, khatarta wasaqowga waa dhexdhexaad.
Xinyi TD1 PCR dhijitaalka ah ee yar yarXinyi TD1 waa madal PCR dhijitaal ah oo gudaha ah, habka ogaanshaha aasaasiga ah: soo saar 30,000-50,000 dhibco biyo-saliid ah iyada oo loo marayo koronto dhaliye, ku xooji qalabka PCR ee caadiga ah, ugu dambeyntiina dhaafo Akhristaha dhibicda ayaa akhriya calaamadda fluorescent ee dhibic kasta.Jiilka dhibicda iyo akhrinta goobtan labadaba waxaa lagu sameeyaa jajab gaar ah oo leh khatarta faddarayntu yar tahay.
STILLA Naica PCR dhijitaalka ah ee yar yarSTILLA Naica waa madal PCR dhijitaal ah oo cusub.Habka lagu ogaanayo aasaasiga ah waa: ku dar xalalka falcelinta ee chip-ka, ku rid chip-ka jiilka yar-yar iyo nidaamka xoojinta, oo soo saar 30,000 micro-dhibcood.Ku faafi chip-ka, iyo kor-u-qaadista PCR-ga ayaa lagu dhammeeyaa chip-ka.Kadibna chip-ka la xoojiyay ayaa loo wareejiyaa nidaamka falanqaynta wax-akhrinta-yar-yar, waxaana calaamadda fluorescent la akhriyaa iyadoo sawirro laga qaadayo.Maadaama hawsha oo dhami ay ku dhacayso jajab xidhan, khatarta faddarayntu way yar tahay.
4. ThermoFisher QuantStudio 3D chip digital PCR
ThermoFisher QuantStudio 3D waa madal kale oo PCR dhijitaalka ah oo chip-ku-saleysan.Nidaamkeeda aasaasiga ah ee lagu ogaanayo waa: ku dar xalalka falcelinta faafiyaha, oo si siman u faafi xalka falcelinta chip-ka oo leh 20,000 microwells iyada oo loo marayo faafiyaha., saar chip-ka mishiinka PCR si aad u xoojiso, oo ugu dambeyntii geli chip-ka akhristaha oo qaado sawir si aad u akhrido calaamadda fluorescent.Qalliinku waa yara adag yahay, habka oo dhanna waxaa lagu fuliyaa jajab xiran, khatarta faddarayntuna waa yar tahay.
5. JN MEDSYS Clarity chip digital PCR
JN MEDSYS caddayntu waa madal PCR dhijitaalka ah oo nooc cusub ah.Nidaamkeeda ogaanshaha aasaasiga ah waa: ku dar xalka falcelinta codsadaha, oo si siman u faafi xalka falcelinta 10,000 tuubooyinka PCR ee ku go'an tuubada PCR iyada oo loo marayo codsadaha.Chip-ka yar-yar, xal-celinta fal-celinta waxay gashaa chip-ka iyada oo loo marayo ficil-celin, iyo tuubada PCR-ga ee chip-ka waxaa lagu dhejiyaa mashiinka PCR si loo kordhiyo, ugu dambeyntiina chip-ka ayaa la gelinayaa akhristaha si uu u akhriyo calaamadda fluorescent adigoo sawir qaadaya.Qalliinku aad ayuu u dhib badan yahay.Khatarta faddarayntu way yar tahay.
Halbeegyada madal kasta oo PCR dhijitaalka ah ayaa loo soo koobay sida soo socota:
Tilmaamayaasha qiimaynta ee madal PCR dhijitaalka ah waa: tirada unugyada kala qaybsan, tirada kanaalada dhaadheer, kakanaanta hawlgalka iyo khatarta wasakhowga.Laakiin waxa ugu muhiimsan waa saxnaanta ogaanshaha.Hal dariiqo oo lagu qiimeeyo aaladaha PCR-ga dhijitaalka ah ayaa ah in la isticmaalo aaladaha PCR-ga dhijitaalka ah ee badan si loo xaqiijiyo midba midka kale, iyo hab kale waa in la isticmaalo walxaha caadiga ah oo leh qiime sax ah.
Faa'iidooyinka dPCR
1.Gaaritaanka tiro dhammaystiran
2.Dareen sare iyo gaar ahaaneed
3.Waxa lagu ogaan karaa shaybaarada koobiga hoose
Khasaaraha dPCR1. Qalab qaali ah iyo reagents 2. Hawlgal adag iyo waqti dheer oo la ogaado
Waqtigan xaadirka ah, saddexda jiil ee tignoolajiyada PCR waxay leeyihiin faa'iidooyin iyo faa'iidooyin u gaar ah, mid walbana wuxuu leeyahay goobo u gaar ah, mana aha xiriir in jiil beddelo kan kale.Horumarka joogtada ah ee tignoolajiyada ayaa ku durtay firfircooni cusub tignoolajiyada PCR, taas oo u sahlaysa in ay furto hal jihada codsiga midba midka kale, taas oo ka dhigaysa in la ogaado aashitada nukliyeerka mid ku haboon oo sax ah.
Xigasho: Dr. Yuan ayaa ku qaadaya tijaabada
Alaabta lagu taliyay:
Waqtiga boostada: Nov-18-2022
