Multiplex qPCR Probe Premix
Saukewa: HCB5051A
TaqMan multiplex qPCR Master Mix (Dye Based) shine riga-kafi don haɓakawa na 2 × na ainihin lokacin ƙididdige PCR wanda ke da girman hankali da ƙayyadaddun ƙayyadaddun, wanda shine shuɗi a launi, kuma yana da tasirin ƙari samfurin.Wannan samfurin shine 2 × Mix pre-mixed reagent wanda ke ba da damar halayen PCR masu kyalli har zuwa huɗu a cikin amsa ɗaya da kyau.Wannan samfurin yana ƙunshe da hanyar rigakafin ƙwayoyin cuta da aka gyaggyara zuwa zafin farawa Taq enzyme, yana haɓaka haɓaka haɓakawa da ƙayyadaddun bayanai.A lokaci guda, wannan samfurin ya inganta haɓaka mai ɗaukar ra'ayi da yawa, wanda zai iya inganta haɓakar haɓakawa da haɓaka ingantaccen haɓakar ƙirar ƙima.Ana iya amfani da wannan samfurin don nazarin ƙididdiga na genotyping da multiplex.
Ƙayyadaddun bayanai
| Zafafan Farawa | Gina-in zafi farawa |
| Hanyar ganowa | Gano na farko-bincike |
| Hanyar PCR | qPCR |
| Polymerase | Taq DNA polymerase |
| Nau'in samfurin | DNA |
Yanayin Ajiya
Ana jigilar samfurin tare da busassun ƙanƙara kuma ana iya adana shi a -25 ~ -15 ℃ na shekara 2.
Umarni
1. AmsaTsari
| Abubuwan da aka gyara | girma (μL) | Ƙarshe Tattaunawa |
| 2× TaqMan multiplex qPCR Master Mix | 12.5 | 1× |
| Haɗin farko (10 μmol/L) a | × | 0.1-0.5 μmol/L |
| Haɗin bincike (10 μmol/L)b | × | 50-250 nmol/L |
| Rox reference rini | 0.5 | 1× |
| Samfurin DNA/cDNA | 1-10 | - |
| ddH2O | har zuwa 25 | - |
Bayanan kula:Mix sosai kafin amfani don guje wa kumfa mai yawa daga girgiza mai ƙarfi.
a.Mahimmanci na farko: Mai haɗawa na farko ya ƙunshi nau'i-nau'i masu yawa, yawanci kowane firamare a matakin ƙarshe na 0.2 μmol/L kuma ana iya daidaita shi tsakanin 0.1 da 0.5 μmol/L kamar yadda ya dace.
b.Matsakaicin bincike: Mix ɗin bincike ya ƙunshi bincike da yawa tare da siginonin haske daban-daban, kuma ana iya daidaita ƙarfin kowane binciken tsakanin 50 zuwa 250 nmol/L bisa ga takamaiman yanayi.
1.Maganar rini na Rox: Ana amfani da shi akan kayan haɓaka PCR na Real Time kamar Applied Biosystems don gyara kuskuren siginar walƙiya da aka haifar tsakanin rijiyoyi;wannan samfurin ba ya ƙunshe da bayanin rini na Rox.Cas #10200 ana bada shawarar idan an buƙata.
2.Tsarin samfuri: qPCR yana da hankali sosai, kuma ana ba da shawarar a tsoma samfurin don amfani.Idan samfuri shine maganin haja na cDNA, girman samfurin bai kamata ya wuce 1/10 na jimlar ƙarar ba.
3.Tsarin amsawa: 25μL, 30μL ko 50 μL ana ba da shawarar don tabbatar da inganci da maimaita ƙarfin haɓakar ƙwayoyin cuta.
4.Shirye-shiryen tsarin: Da fatan za a shirya a cikin babban benci mai tsabta, kuma yi amfani da tukwici da bututun amsawa ba tare da ragowar nuclease ba;ana ba da shawarar yin amfani da tukwici tare da harsashi masu tacewa.Guji gurbacewar giciye da gurɓataccen iska.
2.Shirin maida martani
| Zagayowar mataki | Temp. | Lokaci | Zagaye |
| Na farko-denaturation | 95 ℃ | 5 min | 1 |
| Denaturation | 95 ℃ | dakika 15 | 45 |
| Annealing/Extension | 60 ℃ | 30 seconds |
Bayanan kula:
1.Annealing/Extensewa: Za'a iya daidaita zafin jiki da lokaci daidai gwargwadon ƙimar Tm da aka ƙera.
2.Samun siginar Fluorescence: Lokacin samun siginar walƙiya da ake buƙata don kayan gano qPCR daban-daban ya bambanta, da fatan za a saita gwargwadon ƙayyadaddun ƙayyadaddun lokaci.An saita lokacin kayan aikin gama gari da yawa kamar haka:
Dakika 20: Abubuwan Halittu Masu Amfani 7700, 7900HT, 7500 Mai sauri
31 sec: Applied Biosystems 7300
32 sec: Applied Biosystems 7500
Bayanan kula
Da fatan za a saka PPE da ake buƙata, irin wannan rigar lab da safar hannu, don tabbatar da lafiyar ku da amincin ku!
