Bst 2.0 DNA Polymerase (Glycerol n'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.
Ngwa
| Akụkụ | HC5005A-01 | HC5005A-02 | HC5005A-03 |
| BstDNApolymerase 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.DNA eriri otu mmeghachi omume mgbanwe
3.High GC gene sequencing
4.DNA usoro nke nanogram larịị.
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.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.
3.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.
4.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.
5.E.coli DNA: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
