Dhowr enzymes oo caadi ahaan loo isticmaalo tijaabooyinka PCR

falcelinta silsiladda polymerase, oo loo soo gaabiyo sidaPCRIngiriisi, waa farsamo bayooloji molecular loo isticmaalo in lagu kordhiyo jajabyada DNA-da ee gaarka ah.Waxa loo tixgalin karaa sidii DNA gaar ah oo ku dhufashada jidhka ka baxsan, taas oo si weyn u kordhin karta qadar aad u yar oo DNA ah.Inta lagu jiro oo dhanPCRHabka falcelinta, hal nooc oo walxaha ayaa door muhiim ah ka ciyaara - enzymes.

1. Taq DNA

Tijaabooyinka maalmaha hore eePCR, Saynis yahanadu waxay isticmaaleen Escherichia coli DNA polymerase I, Laakiin waxaa jira dhibaato enzyme this: waxay u baahan tahay in ay buuxiso ensaymes cusub mar kasta oo wareegga la sameeyo, taas oo ka dhigaysa tillaabooyinka qalliinka waxoogaa adag oo ay adagtahay in si buuxda si toos ah loo kordhiyo.Dhibaatadan ayaa la xaliyay ka dib markii saynisyahano si lama filaan ah u go'doomiyeen Taq DNA polymerase ee Thermus aquaticus 1988. Tan iyo markaas, kor u qaadida tooska ah ee DNA-da ayaa noqotay mid dhab ah.Helitaanka ensaymkan ayaa sidoo kale sameeyaPCRtignoolajiyada ku haboon, la taaban karo oo tignoolajiyada caalamiga ah.Hadda, Taq DNA polymerase waa polymerase ugu caansan xirmooyinka DNA-da.

2. PfuDNA

Sida kor ku xusan, Taq DNase waxay leedahay bug weyn, sidaas darteed saynisyahannadu waxay wax ka beddeleen Taq DNA polymerase ilaa xad si ay uga fogaadaan kor u qaadis aan gaar ahayn sababtoo ah isbarbardhigga, taasoo keentay natiijooyin aan sax ahayn.Laakiin wax ka beddelka Taq DNA polymerase wuxuu joojin karaa dhaqdhaqaaqa DNA polymerase heerkulka qolka.PfuDNA polymerase waxay si fiican u hagaajin kartaa faa'iido darrada kor ku xusan ee Taq DNA polymerase, si fal-celinta PCR loo fuliyo si caadi ah, iyo heerka guusha ee xoojinta hiddo-wadaha si wax ku ool ah ayaa loo horumarin karaa.

3. Dib-u-dejinta qoraalka

Reverse transcriptase waxa la helay 1970. Insaymkani waxa uu RNA u adeegsadaa template ahaan, dNTP substrate ahaan, waxa uu raacayaa mabda'a lammaanaynta salka, oo isku xidha DNA hal xadhig ah oo kaabaya qaabka RNA ee jihada 5′-3′.Qoraal-ku-soo-celinta dib-u-celinta waxay ugu horreyn ku xiran tahay waxqabadka DNA-da polymerase ee DNA-da ama qaababka RNA sidaas darteed ma laha 3'-5' dhaqdhaqaaq exonuclease.Si kastaba ha ahaatee, waxay leedahay waxqabadka RNase H, kaas oo xaddidaya dhererka isku-darka qoraalka qoraalka ilaa xad.Sababo la xiriira aaminnimada hoose iyo heerkulbeegga duurjoogta transcriptase, saynisyahannadu sidoo kale wax ka beddeleen.

WaayoPCRTijaabooyinka, waxyaabaha ugu muhiimsan ee la isticmaalo waa: tuubo PCR shaqsiyeed, 4/8-strip PCR tube, PCR taarikada.

LabiyoIsticmaalka PCRleeyihiin kuwan soo socdafaa'iidooyinka:

Taarikada PCR: Iswaafajinta wareegga kulaylka ballaaran;Isbarbardhig heersare ah, oo si fudud loo aqoonsan karo;si fiican u milicsi fluorescence; wanaagsanwareejinta kulaylka;DNase shahaado leh, RNase, DNA, PCR horjoogayaasha, iyo pyrogen-free la tijaabiyay.

Tubooyinka PCR-ga gaarka ah: Uumi u adkaysta; wanaagsanwareejinta kulaylka;Caddayn indhaha oo aad u fiican;DNase shahaado, RNase, DNA, PCR horjoogayaasha, iyo pyrogen-free la tijaabiyay.

4/8-strips tuubooyinka PCR: Darbiyada aadka u dhuuban, caddayn sare; milicsi iftiin wanaagsan;waxay u isticmaali kartaa warshadaha dawooyinka, warshadaha cuntada, iyo bayoolaji molecular; tayo sare leh, walxo PP bikrad ah; Shahaadada DNase, RNase, DNA, PCR horjoogayaasha, iyo la tijaabiyey pyrogen-free.