Mmalite ọkụ Bst 2.0 DNA Polymerase (Glycerol efu)
Bst DNA polymerase V2 sitere na Bacillus stearothermophilus DNA Polymerase I, nke nwere ọrụ 5′→3′ DNA polymerase na ọrụ nnọchi siri ike, mana ọ nweghị ọrụ 5′→3′ exonuclease.Bst DNA Polymerase V2 dabara nke ọma maka mbugharị strand, isothermal amplification LAMP (Agbatị isothermal na-agba agba) na usoro ngwa ngwa.Bst DNA polymerase V2 bụ ụdị mmalite na-ekpo ọkụ dabere na Bst DNA polymerase V2 (HC5005A) nwetara site na teknụzụ mgbanwe mgbanwe, nke nwere ike igbochi ọrụ polymerase DNA n'ime ụlọ okpomọkụ, yabụ enwere ike rụọ ọrụ mmeghachi omume na imepụta ya na okpomọkụ ụlọ iji gbochie ndị na-abụghị -kpọmkwem nkwalite ma melite arụmọrụ mmeghachi omume, na ụdị a nwere ike lyophilized.Tụkwasị na nke ahụ, a na-ahapụ ọrụ ya na okpomọkụ dị elu, n'ihi ya, ọ dịghị mkpa maka nzọụkwụ ntinye dị iche.
Ngwa
| Akụkụ | HC5006A-01 | HC5006A-02 | HC5006A-03 |
Bst DNA polymerase V2 (Glycerol-free) (8U/μL) | 0,2 ml | 1 ml | 10 ml |
| Ihe nchekwa 10 × HC Bst V2 | 1.5 ml | 2 × 1.5 ml | 3 × 10 ml |
| MgSO4(100mM) | 1.5 ml | 2 × 1.5 ml | 2 × 10 ml |
Ngwa
1.LAMP isothermal amplification
2.Ngwongwo DNA nke otu mmeghachi omume mbugharị
3.Usoro mkpụrụ ndụ ihe nketa GC dị elu
4.Usoro DNA nke ọkwa nanogram.
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 25 nmol nke dNTP n'ime ihe na-adịghị edozi acid n'ime nkeji 30 na 65°C.
Njikwa ogo
1.Nnwale ịdị ọcha nke Protein (SDS-PAGE):Ịdị ọcha nke Bst DNA polymerase V2 bụ ≥99% kpebisiri ike site na nyocha SDS-PAGE site na iji nchọpụta Coomassie Blue.
2.EndonucleaseIhe omume:Ntinye nke mmeghachi omume 50 μL nwere opekempe 8 U nke Bst DNA polymerase V2 yana 1 μg λDNA maka awa 16 na 37 ℃ na-arụpụta enweghị mmebi a na-achọpụta dị ka ekpebisiri ike.
3.Ọrụ Exonuclease:Ntinye nke mmeghachi omume 50 μL nwere opekempe 8 U nke Bst DNA polymerase V2 na 1 μg λ -Hind Ⅲ mgbari DNA maka awa 16 na 37 ℃ na-arụpụta enweghị mmebi a na-achọpụta dị ka ekpebisiri ike.
4.Ọrụ Nickase:Ntinye nke mmeghachi omume 50 μL nwere opekempe 8 U nke Bst DNA polymerase V2 nwere 1 μg pBR322 DNA maka awa 16 na 37°C na-arụpụta enweghị mmebi a na-achọpụta dịka ekpebisiri ike.
5.Ọrụ RNase:Ntinye nke mmeghachi omume 50 μL nwere opekempe 8 U nke Bst DNA polymerase V2 yana 1.6 μg MS2 RNA maka awa 16 na 37 Celsius C na-arụpụta enweghị mmebi a na-achọpụta dịka ekpebisiri ike.
6.E. coliDNA:A na-enyocha 120 U nke Bst DNA polymerase V2 maka ọnụnọ nke E. coli genomic DNA na-eji TaqMan qPCR na primers kpọmkwem maka ebe E. coli 16S rRNA.Ihe mmetọ DNA nke E.coli genomic bụ ≤1 Detuo.
Mmeghachi omume LAMP
| Ngwa | 25μL |
| Ihe nchekwa 10 × HC Bst V2 | 2.5 μl |
| MgSO4 (100mM) | 1.5 μl |
| dNTP (10mM nke ọ bụla) | 3.5 μl |
| SYTO™ 16 Green (25×)a | 1.0 μl |
| Ngwakọta mbụb | 6 μl |
| Bst DNA Polymerase V2 (Glycerol-free) (8 U/ul) | 1 μl |
| Ụdị | × μL |
| ddH₂O | Ihe dị ka 25 μl |
ndetu:
1) a.SYTOTM 16 Green (25×): Dị ka mkpa nnwale si dị, a pụrụ iji agba agba ndị ọzọ dochie anya;
2) b.Ngwakọta mbụ: enwetara site na ịgwakọta 20 µ M FIP, 20 µ M BIP, 2.5 µ M F3, 2.5 µ M B3, 5 µ M LF, 5 µ M LB na mpịakọta ndị ọzọ.
Mmeghachi omume na ọnọdụ
1 × HC Bst V2 Buffer, okpomọkụ nnabata dị n'etiti 60°C na 65°C.
Ọkụ anaghị arụ ọrụ
80 Celsius C, 20 nkeji
