2 × HiF Taq da Jagora Mix
Lambar Cat: HCR2014B
HIF Taq tare da Jagora Mix (Tare da Dye) shiri ne don amfani 2 × premixed bayani wanda ya ƙunshi Plus HIF DNA Polymerase, dNTPs, da ingantaccen buffer.Kwayoyin rigakafin monoclonal guda biyu a cikin dakin da zafin jiki wanda ke hana ayyukan polymerase da 3′→ 5′exonuclease ayyukan ana ƙara su zuwa babban mahaɗin don sauƙi da takamaiman takamaiman Hot Start PCR.Ana ƙara haɓakar haɓakar haɓakar maigidan don ba da enzyme tsawon ƙarfin haɓaka juzu'i, tsayin ƙarawa zai iya zama har zuwa 13 kb, enzyme yana da 5′→3′ DNA polymerase aiki da 3′→5′′ Ayyukan exonuclease, amincin sa shine sau 83 na Taq DNA polymerase, wanda shine sau 9 na talakawa DNA polymerase.Ya dace da haɓaka samfuran hadaddun, samfurin haɓakawa shine ƙarshen mara kyau.
2 × HIF Taq tare da Jagora Mix (Tare da Dye) yana da fa'idodi na sauri da sauƙi, babban hankali, ƙayyadaddun ƙaƙƙarfan ƙayyadaddun ƙarfi, kwanciyar hankali mai kyau, da sauransu, tsarin amsawa kawai yana buƙatar ƙara ƙirar ƙira da samfura, kuma ana iya haɓaka su ta biyu- ka'idar mataki, sauƙaƙe matakan gwaji da adana lokaci.Wannan samfurin ya ƙunshi rini mai nuna electrophoresis, kuma samfuran PCR ana iya amfani da su kai tsaye don electrophoresis.Bugu da kari, samfurin kuma ya ƙunshi takamaiman wakili na kariya, ta yadda mahaɗin maigidan zai iya kula da kwanciyar hankali bayan daskarewa da aka maimaita.
Yanayin Ajiya
Ya kamata a adana samfuran a -25 ~ -15 ℃ don 1 shekara.
Ƙayyadaddun bayanai
Ƙayyadaddun samfur | Babban Mix |
Hankali | 2× |
Zafafan Farawa | Wurin Farawa Mai Kyau |
Tsayawa | Batsa |
Gudun amsawa | Mai sauri |
Girman (samfurin ƙarshe) | Har zuwa 13kb |
Sharuɗɗan sufuri | Busasshen ƙanƙara |
Nau'in samfur | Babban amincin PCR premixes |
Umarni
1.Tsarin amsawar PCR
Abubuwan da aka gyara | girma (μL) |
Samfurin DNA | Dace |
Gabatarwa (10 μmol/L) | 2.5 |
Reverse Primer (10 μmol/L) | 2.5 |
2 × HIF Taq da Jagora Mix | 25 |
ddH2O | zuwa 50 |
2.An shawarar yin amfani da samfuri daban-daban
Nau'in samfuri | Ƙara guntu daga 1kb zuwa 10kb |
Genomic DNA | 50-200 NG |
Plasmid ko Viral DNA | 10 pg-20 |
cDNA | 1-2.5 µL (Kada ku wuce 10% na ƙarar amsawar PCR na ƙarshe) |
3.Ƙarfafa Ƙarfafawa
1) Protocol mataki-biyu (samfurin rikitarwa)
Zagayowar mataki | Temp. | Lokaci | Zagaye |
denaturation na farko | 98 ℃ | 3 min | 1 |
Denaturation | 98 ℃ | 10 seconds | 30-35 |
Tsawaita | 68 ℃ | 30 dak/kb | |
Ƙarshe na ƙarshe | 72 ℃ | 5 min | 1 |
2) Protocol mataki uku (ka'ida ta yau da kullun)
Zagayowar mataki | Temp. | Lokaci | Zagaye |
denaturation na farko | 98 ℃ | 3 min | 1 |
Denaturation | 98 ℃ | 10 seconds | 30-35 |
Annealing | 60 ℃ | dakika 20 | |
Tsawaita | 72 ℃ | 30 dak/kb | |
Ƙarshe na ƙarshe | 72 ℃ | 5 min | 1 |
3) Annealing Gradient Protocol (complexity template)
Zagayowar mataki | Zazzabi | Lokaci | Zagaye |
denaturation na farko | 98 ℃ | 3 min | 1 |
Denaturation | 98 ℃ | 10 dakika | 15 (1℃ raguwa a kowane zagaye) |
Rarraba annealing | 70-55 ℃ | dakika 20 | |
Tsawaita | 72 ℃ | 30 dak/kb | |
Denaturation | 98 ℃ | 10 dakika |
20 |
Annealing | 55 ℃ | dakika 20 | |
Tsawaita | 72 ℃ | 30 dak/kb | |
Ƙarshe na ƙarshe | 72 ℃ | 5 min | 1 |
Fasaloli a ƙarƙashin ƙa'idar haɓakawa daban-daban
Protocol | Mataki Biyu | Mataki na uku | Rarraba annealing |
Spec. | sauri | matsakaici | a hankali |
Musamman | babba | matsakaici | babba |
Farashin PCR | matsakaici | babba | matsakaici |
Yawan ganowa | babba | matsakaici | babba |
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!