Mataki Daya RT-qPCR SYBR Green Premix
Saukewa: HCB5140A
Mataki ɗaya RT-qPCR Syber Green Premix shine don ƙididdige ƙimar haske dangane da SYBR Green I rini.Yin amfani da ƙayyadaddun ƙayyadaddun ƙayyadaddun kwayoyin halitta, an kammala rubutun juzu'i da halayen qPCR a cikin bututu guda ɗaya, kawar da buƙatar maimaita buɗewar hula da ayyukan bututu, haɓaka haɓaka haɓakar ƙima da rage haɗarin kamuwa da cuta.Don samfuran RNA, kit ɗin yana amfani da Reverse Transcriptase mai jure zafi don ingantaccen haɗin cDNA da HotStart Taq DNA Polymerase don haɓaka ƙididdigewa.Ƙarƙashin ingantaccen tsarin buffer, azancin kit ɗin zai iya zama sama da 0.1 pg don maƙasudin da aka bayyana sosai kuma har zuwa 1 pg don maƙasudin matsakaicin matsakaici.Kit ɗin ya dace don haɓakawa da ƙididdige samfuran DNA.Yana ba da damar ganowa da ƙididdige adadin acid nucleic daga nau'ikan tsirrai da dabbobi daban-daban, sel da ƙananan ƙwayoyin cuta.
Abubuwan da aka gyara
| No | Suna | Ƙarar | Ƙarar |
| 1 | Babban Buffer | 250 ml | 2 × 1.25 ml |
| 2 | Advanced Enzyme Mix | 20 μl | 200 μl |
| 3 | RNase Free H2O | 250 ml | 2 × 1.25 ml |
Yanayin Ajiya
Ya kamata a adana wannan samfurin a -25 ~ -15 ℃ daga haske har shekara 1.
Umarni
1.Reaction tsarin sanyid
| Abubuwan da aka gyara | girma (μL) | girma (μL) | Natsuwa na ƙarshe |
| Babban Buffer | 12.5 | 25 | 1× |
| Advanced Enzyme Mix | 1 | 2 | - |
| Gabatarwa (10 μmol/L)a | 0.5 | 1 | 0.2 μmol/L |
| Reverse Primer (10 μmol/L)a | 0.5 | 1 | 0.2 μmol/L |
| RNA Tamplateb | X | X | - |
| RNase Free H2Oc | zuwa 25 | zuwa 50 | - |
Bayanan kula:
1) a.TMatsakaicin matakin ƙarshe shine 0.2 μmol/L, wanda kuma za'a iya daidaita shi tsakanin 0.1 da 1μmol/L kamar yadda ya dace.
2) b.Reagent yana da matukar damuwa, tare da Total RNA a cikin kewayon 1pg-1μg, kuma gwajin samfuran ɗan adam ya nuna ingantaccen shigarwar 1 pg-100 ng, yana sarrafa ƙimar Ct gabaɗaya a cikin kewayon 15-30 kamar yadda ya dace.
3) c.Ana ba da shawarar yin amfani da 20μL ko 50μL don tabbatar da inganci da sake haifuwa na haɓaka kwayoyin halitta.
4) d.Da fatan za a shirya a cikin benci mai tsafta kuma yi amfani da tukwici da bututun amsawa mara-kyau;Ana ba da shawarar tukwici tare da harsashi masu tacewa.Guji gurbacewar giciye da gurɓataccen iska.
2.Shirin maida martani
| Zagayowar mataki | Temp. | Lokaci | Zagaye |
| Juya rubutun | 50 ℃a | 6 min | 1 |
| denaturation na farko | 95 ℃ | 5 min | 1 |
| Amplification dauki | 95 ℃ | dakika 15 | 40 |
| 60 ℃b | 30 seconds | ||
| Narkar da lankwasa mataki | Defaults na Kayan aiki | 1 | |
Bayanan kula:
1) a.Za'a iya zaɓar yanayin jujjuyawar juyi tsakanin 50-55°C bisa ga buƙatun gwaji.Don samfuran DNA, ana iya barin tsarin rubutun baya.
2) b.A cikin lokuta na musamman za'a iya daidaita zafin jiki na annealing/tsawo bisa ga ƙimar Tm na farko, ana bada shawarar 60°C.
Bayanan kula
1. Wannan samfurin don amfanin bincike ne kawai.
2. Da fatan za a yi aiki da riguna na lab da safofin hannu masu yuwuwa, don amincin ku.
