2× Degdeg ah Taq Super Mix
Bisadda No: HCR2016A
2× Degdegga Taq Super Mix waxay ku salaysan tahay Taq DNA Polymerase oo wax laga beddelay, iyadoo lagu darayo arrin fidineed oo xooggan, qodobka kobcinta iyo hab-dhisidda la wanaajiyey, oo leh hufnaan sare oo sarreeya.Xawaaraha balaadhinta qaab-dhismeedka kakan sida genome gudaha 3kb waxa uu gaadhaa 1-3 sek/kb, iyo ta qaab-dhismeedka fudud sida plasmids gudaha 5kb waxa uu gaadhaa 1 sek/kb.Alaabtani waxay si weyn u badbaadin kartaa waqtiga falcelinta PCR.Isla mar ahaantaana, isku-darka ayaa ka kooban dNTP iyo Mg2+, kuwaas oo lagu kordhin karo kaliya iyada oo lagu darayo furayaasha iyo qaababka, taas oo sidoo kale si weyn u fududaynaysa tallaabooyinka hawlgalka ee tijaabada.Intaa waxaa dheer, isku dhafka ah wuxuu ka kooban yahay dheeha elektrophoretic, kaas oo si toos ah u noqon kara electrophoresis ka dib falcelinta.Wakiilka difaaca ee alaabtan ayaa ka dhigaysa isku darka inuu ilaaliyo dhaqdhaqaaqa deggan ka dib marka la qaboojiyo soo noqnoqda iyo dhalaalidda.Qaybta 3'-dhamaadka A ee alaabta PCR si fudud ayaa loogu xidhi karaa qaybta T.
Qaybaha
2× Degdeg ah Taq Super Mix
Xaaladaha Kaydinta
Alaabta PCR Master Mix waa in lagu kaydiyaa -25~-15℃ ilaa 2 sano.
Tilmaamaha
| Tilmaamaha alaabta | Degdeg ah Taq Super Mix |
| Xoog-saarid | 2× |
| Bilow kulul | Ku-dhismay Hot Start |
| Dulqaadasho | 3′-A |
| Xawaaraha falcelinta | Degdeg ah |
| Baaxadda (Badeecada u dambaysa) | Ilaa 15 kb |
| Shuruudaha gaadiidka | Baraf qallalan |
Tilmaamaha
1. Nidaamka falcelinta (50 μL)
| Qaybaha | Cabbirka (μL) |
| Qaabka DNA* | ku habboon |
| Qoryaha hore ee hore (10 μmol/L) | 2.5 |
| Dib-u-celinta hore (10 μmol/L) | 2.5 |
| 2× Degdeg ah Taq Super Mix | 25 |
| ddH2O | ilaa 50 |
2.Kordhinta Hab-maamuuska
| Tallaabooyinka wareegga | Heerkulka (°C) | Waqtiga | Wareegtada |
| Predenaturation | 94 | 3 daq | 1 |
| Denaturation | 94 | 10 ilbiriqsi |
28-35 |
| Annealing | 60 | 20 ilbiriqsi | |
| Kordhinta | 72 | 1-10 ilbiriqsi/kb |
Isticmaalka qaabab kala duwan oo lagu taliyay:
| Nooca template | Qaybta isticmaalka (nidaamka falcelinta 50 μL) |
| DNA-da genomic ama dareeraha E. coli | 10-1,000 ng |
| Plasmid ama DNA viral | 0.5-50 ng |
| CDNA | 1-5 µL (aan ka badnayn 1/10 ee mugga guud ee falcelinta PCR) |
| Lagu taliyay isticmaalka habab kala duwan | |
Xusuusin:
1.Isticmaalka reagent: si buuxda u dhalaali oo isku qas ka hor isticmaalka.
2. Heerkulka xajinta: Heerkulka annealing waa qiimaha caalamiga ah ee Tm, waxaana sidoo kale lagu dejin karaa 1-2℃ ka hooseeya qiimaha Tm ee asaasiga ah.
3. Xawaaraha fidinta: U deji 1 sek/kb qaabab kakan sida genome iyo E. coli gudaha 1 kb;u dhig 3 ilbiriqsi/kb qaabab isku dhafan sida 1-3 kb genome iyo E. coli;u dhig 10 ilbiriqsi/kb qaab-dhismeedka adag ee ka badan 3 kb genome iyo E. coli.Waxaad u dejin kartaa qiimaha 1 ilbiriqsi/kb qaab fudud sida plasmid ka yar 5 kb, 5 sek/kb qaab fudud sida plasmid inta u dhaxaysa 5 iyo 10 kb, iyo 10 ilbiriqsi/kb qaab fudud sida plasmid ka weyn 10 kb.
Xusuusin
1. Badbaadadaada iyo caafimaadkaaga, fadlan xidho jaakadaha shaybaadhka iyo galoofyada la tuuri karo si loogu hawlgalo.
2. Alaabtan waxaa loogu talagalay isticmaalka cilmi-baarista oo keliya!
