Rubutun juzu'i na al'ada ba zai iya jure yanayin zafi mai girma ba (mafi kyawun zafin jiki don ayyukan MMLV shine 37-50°C, kuma AMV shine 42-60°C).RNA mafi hadaddun ƙwayar cuta ba za a iya jujjuya shi yadda ya kamata zuwa cDNA a ƙananan zafin jiki ba, yana haifar da ingantaccen ganowa Ragewar.RT-qPCR na al'ada gabaɗaya yana buƙatar shigar da mahimman enzymes guda biyu (reverse transcriptase da DNA polymerase), wanda ke sa ba zai yiwu a sauƙaƙe aikin tsarin amsawa da rage farashi ba.Anan zamu gabatar da manyan juzu'i biyu masu jure zafin jiki, TtH da RevTaq.Wadannan enzymes guda biyu kuma suna da aikin DNA polymerase, don haka ana kiran su bifunctional enzymes.

TTH DNA polymerase

Dole ne ku ji labarin TtH, wanda aka samo shi daga kwayoyin thermophilic Thermus thermophilus HB8.A gaban divalent cations kamar Mg2+, yana da DNA polymerase aiki.Ana amfani dashi sosai a cikin halayen PCR kamar Taq enzyme, amma yana da juriya mai zafi fiye da Taq enzyme, don haka yana da tasiri mafi kyau akan PCR tare da samfuran abun ciki na GC masu girma.

Wannan enzyme m ba shi da 3′→5′ exonuclease ayyuka da 5′→3′ exonuclease ayyuka, don haka shi kuma za a iya amfani da dideoxy sequencing.

Wannan enzyme yana da aikin RTase.A gaban Mn2+, za a inganta ayyukan RTase.Yin amfani da wannan fasalin, ana iya amfani da shi don aiwatar da martanin juzu'i da amsawar PCR a cikin bututu ɗaya, wato, RT-PCR-mataki ɗaya.Koyaya, a gaban Mn2+, daidaiton RT-PCR bai yi girma ba.Ayyukan RT ba shi da alaƙa da ayyukan rnaase H.

Ƙara yawan aiki na Tth-DNA polymerase (pH9, mafi girma +55 ℃～ + 70 ℃, matsakaicin + 95 ℃) yana shawo kan matsalolin da tsarin sakandare na RNA ya haifar.Sakamakon cDNA na iya haɓaka ta PCR tare da enzyme iri ɗaya a gaban Mg2+ ions.

Ƙarfin Tth-DNA polymerase don yin juzu'in juzu'i da haɓaka DNA a yanayin zafi mai girma yana sa wannan enzyme yana da amfani ga ƙididdiga na RT-PCR, cloning, da nazarin maganganun kwayoyin halitta na salon salula da kwayar cutar RNA.
Ana amfani da polymerase Tth-DNA don RT-PCR don haɓaka RNA har zuwa 1kb.

Features da abũbuwan amfãni

DNA polymerase:

• Tabbatar da ingantacciyar sarkar polymerase (PCR) girman samfurin, aƙalla 1000 bp a cikin martanin RT-PCR.

• Karɓar gyare-gyaren deoxyribonucleoside triphosphate a matsayin maƙalli

Ba shi da alaƙa da ayyukan RNase H

• Yana da babban kwanciyar hankali na thermal don shawo kan matsaloli, yawanci yana da alaƙa da babban tsarin sakandare da ke cikin RNA

RevTaq RT-PCR DNA polymerase

DNA polymerase mai jurewa zafi tare da aikin jujjuyawar rubutu

RevTaq-RT-PCR-DNA polymerase injiniyan injiniya ne, mai juriya mai zafi, mai aiki dual-aiki tare da juzu'i na juzu'i da ayyukan DNA polymerase da aka samu ta hanyar jagora da juyin halitta.

Rabin rayuwar RevTaq RT-PCR DNA polymerase a 95°C ya fi mintuna 40 girma.

RevTaq RT-PCR DNA polymerase yana ba da damar jujjuya yanayin zafi kai tsaye daga samfurin RNA, kuma ana iya maimaita matakin jujjuyawar sau da yawa don samar da ƙarin samfuran cDNA.

RevTaq RT-PCR DNA polymerase yana ba da damar “mataki-sifili” RT-PCR (babu matakin jujjuyawar isothermal), saboda a cikin matakin tsawo na PCR na cyclic, juzu'i da haɓaka DNA suna faruwa lokaci guda.Wannan kuma yana haɓaka martanin juzu'i a yanayin zafi mai girma, ta haka yana rage matsalolin da ake fuskanta wajen narkar da ƙaƙƙarfan tsarin na biyu a cikin RNA a babban yanayin zafi.

Saboda dabarar farawa mai zafi na tushen aptamer, RevTaq RT-PCR DNA polymerase zai samar da sakamako mai kyau lokacin da zafin jiki na annealing da tsawo ya wuce 57°C.

Tunda enzyme yana da tsayayya da zafi, ana bada shawara don ƙirƙira ƙirar ƙira da bincike tare da manyan abubuwan narkewa (> 60 ° C).

Ana ba da shawarar inganta yanayin zafi na matakin cirewa/tsawo ta hanyar rage zafin jiki yayin aiwatar da saitin amsawa.

Mafi girman zafin jiki, mafi girman ƙayyadaddun PCR.Juya juzu'i yawanci ana yin shi ne a mafi girma fiye da zagayowar PCR, saboda DNA Primer: RNA Template hybridization yawanci yana da wurin narkewa sama da DNA Primer: cDNA Template duplex.

RevTaq RT-PCR DNA polymerase an tsara shi ta hanyar halitta kuma an inganta shi, kuma girman amplicon yana tsakanin 60-300 bp.

RevTaq RT-PCR DNA polymerase iyakar ganowa ya kai kwafi 4/

An nuna ingantaccen tsarin amsawa (kafa manyan maƙallan narkewa) a cikin adadi.RevTaq RT-PCR DNA polymerase-driven RT-PCR yana nuna mafi kyawun hankali fiye da TaqPath 1-mataki RT-qPCR babban mahaɗin, da ƙananan ganewa Samfurin dilution gradient.

Ƙarin fa'idodi:

Ayyukan farawa da sauri → Za a iya tsallake matakin haƙar zafi na farko.

Tsarin aptamer mai zafi-farawa → Samar da aikin enzyme 100% nan da nan kuma ya hana haɓaka da ba takamaiman ba a ƙananan yanayin zafi (<57°C).

Aikin share fage → An cire matakin cirewar RNA, saboda RevTaq RT-PCR DNA polymerase shima yana iya aiwatar da samfuran halayen ɗan adam.Nan da nan zai iya lalata membranes tantanin halitta na eukaryotes, ƙwayoyin cuta da ƙwayoyin cuta a cikin zagayowar RT-PCR mai zafi.

IVD albarkatun kasa matakin → high quality matsayin da kuma musamman m farashin