Hotstart Taq DNA Polymerase
Hot Start Taq DNA Polymerase (Mgbanwe Antibody) bụ DNA polymerase na-ekpo ọkụ na-ekpo ọkụ sitere na Thermus aquaticus YT-1, nke nwere ọrụ 5′→3′ polymerase na ọrụ 5′ flap endonuclease.Ihe mmalite Taq DNA polymerase na-ekpo ọkụ bụ Taq DNA polymerase nke ọgwụ mgbochi Taq thermolabile gbanwere.Mgbanwe mgbochi mmadụ mụbara nkọwapụta, nghọta, na mkpụrụ nke PCR.
Ngwa
| Akụkụ | HC1012A-01 | HC1012A-02 | HC1012A-03 | HC1012A-04 |
| 5 × HC Taq Buffer | 4 × 1 ml | 4 × 10 ml | 4 × 50 ml | 5 × 400 ml |
| Hot Start Taq DNA Polymerase (Antibody gbanwetụrụ) (5 U/μL) | 0,1 ml | 1 ml | 5 ml | 10 × 5 ml |
Ngwa
10 mM Tris-HCl (pH 7.4 na 25 ℃), 100 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 0.5% Tween20, 0.5% IGEPALCA-630 na 50% Glycerol.
Ọnọdụ Nchekwa
Ụgbọ njem n'okpuru 0 Celsius ma debe ya na -25C ~ -15C.
Nkọwa nkeji
A kọwapụtara otu nkeji dị ka ọnụọgụ enzyme nke na-etinye 15 nmol nke dNTP n'ime ihe na-adịghị edozi acid n'ime nkeji 30 na 75°C.
Njikwa ogo
1.Endonuclease Ọrụ:Ntinye nke 20 U nke enzyme na 4 μg pUC19 DNA maka awa 4 na 37 ℃ arụpụtaghị mmebi nke DNA nke nwere ike ịchọpụta dị ka gel electrophoresis siri kpebie.
2.5kb Lambda PCR:Usoro 25 nke nkwalite PCR nke 5 ng Lambda DNA nwere nkeji 1.25 nke Taq DNA Polymerase n'ihu 200 µM dNTPs na 0.2 µM primers na-arụpụta ngwaahịa 5 kb a na-atụ anya ya.
3.Ọrụ Exonuclease:Ntinye nke mmeghachi omume 50 µl nwere opekempe 12.5 U nke Taq DNA Polymerase nwere 10 nmol 5′-FAM oligonucleotide maka nkeji 30 na 37℃ anaghị ewepụta nbibi nke achọpụtara.
4.Ọrụ RNase:Mweghachi nke mmeghachi omume 10 µL nwere 20 U nke enzyme nwere 1μg nke transcript RNA maka awa 2 na 37 Celsius arụpụtaghị mmebi nke RNA enweghị ike ịchọpụta dị ka gel electrophoresis siri kpebie.
5.Agbanye ọkụ:Mba.
Sistemụ mmeghachi omume
| Ngwa | Olu |
| DNA templatea | nhọrọ |
| 10 μM na-aga n'ihu | 0,5 μl |
| 10 μM Reverse Primer | 0,5 μl |
| Ngwakọta dNTP (10mM nke ọ bụla) | 0,5 μl |
| 5 × HC Taq Buffer | 5 μl |
| DNA Polymeraseb(5U/μL) | 0.125 μl |
| Mmiri na-enweghị nuklia | Ihe dị ka 25 μl |
ndetu:
1) a.
| DNA | Ọnụ ego |
| Genomic | 1ng-1 μg |
| Plasmid ma ọ bụ Viral | 1 pg-1 nkp |
2) b.Nleta kacha mma nke Taq DNA Polymerase nwere ike ịdị site na 5-50 nkeji/mL (0.1-0.5 nkeji/25 µL mmeghachi omume) na ngwa pụrụ iche.
Usoro ịgba ígwè okpomọkụ
PCR
| Nzọụkwụ | Okpomọkụ(Celsius C) | Oge | okirikiri |
| denaturation nke mbụa | 95 ℃ | 1-3 nkeji | - |
| Denaturation | 95 ℃ | 15-30 nkeji | Usoro 30-35 |
| Na-ewe iweb | 45-68 ℃ | 15-60 nkeji | |
| Mgbatị | 68 ℃ | 1kb/min | |
| Mgbatị ikpeazụ | 68 ℃ | 5mins | - |
ndetu:
1) Mbipụta mbụ nke 1 min na 95°C zuru oke maka ọtụtụ nkwalite.Maka ndebiri ndị siri ike, a na-atụ aro ogologo denaturation nke 2-3mins na 95°C.Site na PCR colony, a na-akwado denaturation 5mins mbụ na 95 Celsius.
2) The annealing nzọụkwụ bụ 15-60 s.Annealing okpomọkụ dabere na Tm nke primer ụzọ na-emekarị 45-68 ℃.
