Wild Taq DNA Polymerase
ʻO Taq DNA Polymerase kahi DNA polymerase thermostable mai Thermus aquaticus YT-1, nona ka hana polymerase 5′→3′ a me ka hana 5′ flap endonuclease.
Nā ʻāpana
| ʻāpana | HC1010A-01 | HC1010A-02 | HC1010A-03 | HC1010A-04 |
| 10× Taq Buffer | 2×1 mL | 2×10 mL | 2×50 mL | 5×200 mL |
| Taq DNA Polymerase (5 U/μL) | 0.1 mL | 1 mL | 5 mL | 5×10 mL |
Kūlana Waihona
ʻO ka halihali ma lalo o 0°C a mālama ʻia ma -25°C~-15°C.
Wehewehe Unite
Hoʻokahi ʻāpana i wehewehe ʻia ʻo ka nui o ka enzyme e hoʻokomo i 15 nmol o dNTP i loko o ka mea hiki ke hoʻoheheʻe ʻia i loko o 30 mau minuke ma 75°C.
Ka Mana Mana
1.Hoʻomaʻemaʻe Protein (SDS-PAGE):ʻO ka maʻemaʻe o Taq DNA polymerase he ≥95% i hoʻoholo ʻia e ka SDS-PAGE analysis.
2.Endonuclease Activity:ʻO ka liʻiliʻi loa o 5 U o Taq DNA polymerase me 1 μg λDNA no 16 mau hola ma 37 ℃ ka hopena ʻaʻole hiki ke ʻike ʻia e like me ka mea i hoʻoholo ʻia.
3.Ka hana Exonuclease:ʻO ka liʻiliʻi loa o 5 U o Taq DNA polymerase me 1 μg λ -Hind Ⅲ digest DNA no 16 mau hola ma 37 ℃ hopena ʻaʻole hiki ke ʻike ʻia ka degradation e like me ka mea i hoʻoholo ʻia.
4.Hana Nickase:ʻO ka liʻiliʻi o 5 U o Taq DNA polymerase me 1 μg pBR322 DNA no 16 mau hola ma 37°C ʻaʻohe hopena i ʻike ʻia e like me ka mea i hoʻoholo ʻia.
5.Hana Hana:ʻO ka liʻiliʻi o 5 U o Taq DNA polymerase me 1.6 μg MS2 RNA no 16 mau hola ma 37°C ʻaʻohe hopena i ʻike ʻia e like me ka mea i hoʻoholo ʻia.
6.E. coliDNA:5 U o Taq DNA polymerase ua hoʻopaʻa ʻia no ka loaʻa ʻana o E. coli genomic DNA me ka hoʻohana ʻana i TaqMan qPCR me nā primers kikoʻī no ka E. coli 16S rRNA locus.ʻO ka E. coli genomic DNA contamination he ≤1 Kope.
7.Hoʻonui PCR (5.0 kb Lambda DNA)- He hopena 50 µL i loaʻa iā 5 ng Lambda DNA me 5 mau ʻāpana o Taq DNA Polymerase no 25 mau pōʻai o ka hoʻonui ʻana i ka PCR i ka huahana 5.0 kb i manaʻo ʻia.
Hoʻonohonoho ʻana
| Nā ʻāpana | Volume |
| DNA templatea | koho |
| 10 μM Puhi mua | 1 μL |
| 10 μM Pumi Huli | 1 μL |
| Hui dNTP (10mM kēlā me kēia) | 1 μL |
| 10×Taq Buffer | 5 μL |
| Taq DNA Polymeraseb | 0.25 μL |
| Wai nuclease-free | A hiki i 50 μL |
Nā memo:
1) He ʻokoʻa ka manaʻo o ka hopena maikaʻi loa o nā mamana like ʻole.Hōʻike ka papa ma lalo nei i ka hoʻohana ʻana i ka laʻana i manaʻo ʻia o 50 µL ʻōnaehana pane.
| DNA | Ka nui |
| Genomic | 1 ng-1 μg |
| Plasmid a i ʻole Viral | 1 pg-1 ng |
2) ʻO ka manaʻo maikaʻi loa o Taq DNA Polymerase mai ka 0.25 µL~1 µL i nā noi kūikawā.
PanePapahana
| ʻanuʻu | Mahana(°C) | Manawa | Nā pōʻaiapuni |
| Denaturation muaa | 95 ℃ | 5 min | - |
| Denaturation | 95 ℃ | 15-30 s | 30-35 Puka |
| Hoʻopilib | 60 ℃ | 15 s | |
| Hoʻonui | 72 ℃ | 1kb/min | |
| Hoʻonui hope | 72 ℃ | 5 min | - |
Nā memo:
1) Ua kūpono ke kūlana denaturation mua no ka hapa nui o nā hopena amplification a hiki ke hoʻololi ʻia e like me ka paʻakikī o ka hoʻolālā template.Inā paʻakikī ka hoʻolālā template, hiki ke hoʻonui ʻia ka manawa pre-denaturation i 5 - 10mins e hoʻomaikaʻi i ka hopena denaturation mua.
2) Pono e hoʻoponopono ʻia ka mahana annealing e like me ka waiwai Tm o ke kumu mua, i hoʻonohonoho maʻamau ʻia i 3 ~ 5 ℃ haʻahaʻa ma mua o ka waiwai Tm o ka primer;No nā mamana paʻakikī, pono e hoʻoponopono i ka mahana annealing a hoʻolōʻihi i ka manawa hoʻonui e hoʻokō pono i ka amplification.
