Hotstart Taq DNA Polymerase
Hot Start Taq DNA Polymerase (gyaran Antibody) shine farkon thermostable DNA polymerase daga Thermus aquaticus YT-1, wanda ke da aikin 5′→3′ polymerase da aikin 5′ flap endonuclease.Zazzafan farko Taq DNA polymerase shine Taq DNA polymerase wanda aka gyara ta antibodies Taq thermolabile.Canjin rigakafin ƙwayar cuta ya ƙaru ƙayyadaddun ƙayyadaddun, hankali, da yawan amfanin PCR.
Abubuwan da aka gyara
| Bangaren | HC1012 A-01 | HC1012 A-02 | HC1012 A-03 | HC1012 A-04 |
| 5 × HC Taq Buffer | 4 × 1 ml | 4 × 10 ml | 4 × 50 ml | 5 × 400 ml |
| Hot Start Taq DNA Polymerase (Antibody modified) (5 U/μL) | 0.1 ml | 1 ml | ml 5 | 10 × 5 ml |
Aikace-aikace
10 mM Tris-HCl (pH 7.4 a 25 ℃), 100 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 0.5% Tween20, 0.5% IGEPALCA-630 da 50% Glycerol.
Yanayin Ajiya
Jirgin da ke ƙarƙashin 0 ° C kuma a adana shi a -25 ° C ~ -15 ° C.
Ma'anar Naúrar
An ayyana raka'a ɗaya azaman adadin enzyme wanda ya haɗa 15 nmol na dNTP cikin abu maras narkewa a cikin mintuna 30 a 75°C.
Kula da inganci
1.EndAyyukan onuclease:Ƙunƙarar 20 U na enzyme tare da 4 μg pUC19 DNA na 4 hours a 37 ℃ ya haifar da rashin lahani na DNA kamar yadda gel electrophoresis ya ƙaddara.
2.5kb Lambda PCR:25 Zagaye na haɓaka PCR na 5 ng Lambda DNA tare da raka'a 1.25 na Taq DNA Polymerase a gaban 200 µM dNTPs da 0.2 µM na farko yana haifar da samfurin 5 kb da ake tsammanin.
3.Ayyukan Exonuclease:Haɓaka amsawar 50 µl mai ɗauke da ƙaramar 12.5 U na Taq DNA Polymerase tare da 10 nmol 5′-FAM oligonucleotide na mintuna 30 a 37℃ yana haifar da rashin lahani da za a iya ganowa.
4.Ayyukan RNase:Haɓaka halayen 10 µL mai ɗauke da 20 U na enzyme tare da 1μg na kwafin RNA na awanni 2 a 37°C ya haifar da rashin lahani na RNA kamar yadda gel electrophoresis ya ƙaddara.
5.Rashin kunna zafi:A'a.
Tsarin martani
| Abubuwan da aka gyara | Ƙarar |
| Samfurin DNAa | na zaɓi |
| 10 μM Gabatarwa | 0.5 μl |
| 10 μM Reverse Primer | 0.5 μl |
| dNTP Mix (10mM kowane) | 0.5 μl |
| 5 × HC Taq Buffer | 5 μl |
| Taq DNA Polymeraseb(5U/μL) | 0.125 ml |
| Ruwa mara nukiliya | Har zuwa 25 μl |
Bayanan kula:
1) a.
| DNA | Adadin |
| Genomic | 1 ng-1 μg |
| Plasmid ko Viral | 1 shafi-1 ng |
2) b.Mafi kyawun maida hankali na Taq DNA Polymerase na iya zuwa daga raka'a 5-50/mL (0.1-0.5 raka'a/25 µL amsa) a cikin aikace-aikace na musamman.
Ƙa'idar hawan keke na thermal
PCR
| Mataki | Zazzabi(°C) | Lokaci | Zagaye |
| denaturation na farkoa | 95 ℃ | 1-3 min | - |
| Denaturation | 95 ℃ | 15-30 s | Zagaye 30-35 |
| Annealingb | 45-68 ℃ | 15-60s | |
| Tsawaita | 68 ℃ | 1kb/min | |
| Ƙarshe Ƙarshe | 68 ℃ | 5 min | - |
Bayanan kula:
1) Ƙimar farko na 1 min a 95 ° C ya wadatar don yawancin haɓakawa.Don samfura masu wahala, ana ba da shawarar tsawan tsayin daka na 2-3mins a 95°C.Tare da PCR na mulkin mallaka, ana ba da shawarar rarrabuwar farko na 5mins a 95°C.
2) Annealing mataki ne yawanci 15-60 s.Matsakaicin zafin jiki ya dogara ne akan Tm na nau'in farko kuma yawanci shine 45-68 ℃.
