Multiplex qPCR Probe Premix
Bisadda No: HCB5051A
TaqMan multiplex qPCR Master Mix (Dye Based) waa xal horudhac ah oo loogu talagalay 2 × waqtiga-dhabta ah ee cabbiraadda PCR-ga oo lagu garto dareen sare iyo gaar ahaan, kaas oo midab buluug ah leh, oo leh saamaynta muunadda isku-darka.Alaabtani waa 2× Mix horay isku darka reagent kaas oo awood u siinaya ilaa afar fal-celinta PCR ee fluorescent ee hal falcelin si fiican.Alaabtani waxa ay ka kooban tahay habka antibody-ka ee hidde ahaan wax laga beddelay si loo bilaabo-kuleelka Taq enzyme, oo si weyn u wanaajinaya dareenka xoojinta iyo gaarnimada.Isla mar ahaantaana, sheygani wuxuu si qoto dheer u hagaajiyay bakhti-celinta falcelinta badan, taas oo hagaajin karta waxtarka xoojinta falcelinta iyo kor u qaadida wax-ku-oolka wax-ku-oolka ah ee qaababka-fiirsashada hoose.Alaabtan waxaa loo isticmaali karaa genotyping iyo falanqaynta tirada badan.
Tilmaamid
| Bilow kulul | Bilawga kulul ee la dhisay |
| Habka ogaanshaha | Ogaanshaha hore ee baaritaanka |
| Habka PCR | qPCR |
| Polymerase | Taq DNA polymerase |
| Nooca muunada | DNA |
Xaaladaha Kaydinta
Alaabta waxaa lagu raray baraf qallalan waxaana lagu kaydin karaa -25 ~ -15℃ 2 sano.
Tilmaamaha
1. FalcelintaNidaamka
| Qaybaha | Mugga (μL) | U-fiirsashada kama dambaysta ah |
| 2× TaqMan multiplex qPCR Master Mix | 12.5 | 1× |
| Isku darka koowaad (10 μmol/L) a | × | 0.1 - 0.5 μmol/L |
| Isku darka baaritaanka (10 μmol/L)b | × | 50 - 250 nmol/L |
| Rox tixraaca dheeha | 0.5 | 1× |
| Qaabka DNA/cDNA | 1-10 | - |
| ddH2O | ilaa 25 | - |
Xusuusin:Si fiican isku qas ka hor intaanad isticmaalin si aad uga fogaato xumbo xad dhaaf ah oo ka yimaada gariir xoog leh.
a.Feejignaanta koowaad: Isku-darka Primer wuxuu ka kooban yahay lammaane badan oo asal ah, badiyaa mid kasta oo curiye ah marka ugu dambeysa ee uruurinta 0.2 μmol/L sidoo kale waxaa lagu hagaajin karaa inta u dhaxaysa 0.1 iyo 0.5 μmol/L sida ugu habboon.
b.Diiradda baaritaanka: Isku-dhafka baaritaanka wuxuu ka kooban yahay baarayaal badan oo leh calaamado dhaadheer oo kala duwan, iyo xoogga saarista baaritaan kasta waxaa lagu hagaajin karaa inta u dhaxaysa 50 iyo 250 nmol/L iyadoo loo eegayo xaalad gaar ah.
1.Tixraaca dheeha Rox:Waxa loo istcimaalaa aaladda kor u qaadida PCR ee wakhtiga-dhabta ah sida hab-nololeedka la dabaqay si loo saxo khaladka calaamada iftiinka iftiinka ee ceelasha dhexdooda;Alaabtaan kuma jiraan tixraaca dheeha Rox.Cas#10200 ayaa lagula talinayaa haddii loo baahdo.
2.Qaababka qaabaysan: qPCR aad bay xasaasi u tahay, waxaana lagu talinayaa in la milo template si loo isticmaalo.Haddii qaabku yahay xalka kaydka cDNA, mugga template waa inaanu ka badnayn 1/10 ee mugga guud.
3.Habka falcelinta: 25μL, 30μL ama 50 μL ayaa lagula talinayaa si loo hubiyo waxtarka iyo soo noqnoqda ee kordhinta hidda-wadaha.
4.Diyaarinta nidaamka: Fadlan ku diyaari kursiga kaydka ah ee nadiifka ah, oo isticmaal talooyinka iyo tuubooyinka falcelinta iyada oo aan hadhaaga nukliyeerka;waxaa lagu talinayaa in la isticmaalo talooyinka leh kartoonada shaandhada.Ka fogow wasakhowga isdhaafka ah iyo wasakhowga hawada.
2.Barnaamijka falcelinta
| Tallaabo wareeg | Heerkulka. | Waqtiga | Wareegtada |
| Bilowga-denaturation | 95 ℃ | 5 daqiiqo | 1 |
| Denaturation | 95 ℃ | 15 ilbiriqsi | 45 |
| Baabi'inta/kordhinta | 60 ℃ | 30 ilbiriqsi |
Xusuusin:
1.Annealing/Extension: Heerkulka iyo wakhtiga ayaa si ku habboon loo hagaajin karaa iyadoo loo eegayo qiimaha Tm ee loo qaabeeyey.
2.Helitaanka signalka fluorescence: Wakhtiga helista signalka fluorescence ee looga baahan yahay qalabyada kala duwan ee qPCR ogaanshaha waa ka duwan yahay, fadlan u dhig si waafaqsan wakhtiga ugu yar.Waqtiga dhowr qalab oo caadi ah ayaa loo dejiyay sida soo socota:
20 ilbiriqsi: Nidaamyada noolaha ee la dabaqay 7700, 7900HT, 7500 Degdeg ah
31 ilbiriqsi: Biosystems Applied 7300
32 ilbiriqsi: Nidaamyada noolaha ee la dabaqay 7500
Xusuusin
Fadlan xidho PPE-ga lagama maarmaanka ah, sida jaakada shaybaadhka iyo galoofyada, si loo hubiyo caafimaadkaaga iyo badbaadadaada!
