Mofuthu oa ho Qala Bst 2.0 DNA Polymerase (Glycerol mahala)
Bst DNA polymerase V2 e tsoa ho Bacillus stearothermophilus DNA Polymerase I, e nang le ts'ebetso ea 5′→3' DNA polymerase le ts'ebetso e matla ea ketane, empa ha ho na 5′→3′ exonuclease.Bst DNA Polymerase V2 e loketse hantle bakeng sa ho falla ha strand, isothermal amplification LAMP (Loop mediated isothermal amplification) le tatellano e potlakileng.Bst DNA polymerase V2 ke mofuta oa ho qala o chesang o thehiloeng ho Bst DNA polymerase V2 (HC5005A) e fumanoeng ke theknoloji e fetolehang, e ka thibelang ts'ebetso ea DNA polymerase ka mocheso oa kamore, kahoo sistimi ea karabelo e ka sebetsoa le ho etsoa ka mocheso oa kamore ho thibela ho se sebetse. - ho matlafatsa ka ho khetheha le ho ntlafatsa katleho ea karabelo, 'me phetolelo ena e ka ba lyophilized.Ho phaella moo, mosebetsi oa eona o lokolloa ka mocheso o phahameng, kahoo ha ho hlokahale hore ho be le mohato o fapaneng oa ts'ebetso.
Likaroloana
| Karolo | HC5006A-01 | HC5006A-02 | HC5006A-03 |
Bst DNA polymerase V2 (e se nang Glycerol) (8U/μL) | 0.2 mL | 1 mL | 10 ml |
| 10 × HC Bst V2 Buffer | 1.5 mL | 2×1.5 mL | 3 × 10 mL |
| MgSO4(100mM) | 1.5 mL | 2×1.5 mL | 2 × 10 mL |
Lisebelisoa
1.Ntlafatso ea motlakase ea LAMP
2.DNA strand single displacement reaction
3.Tatelano ea liphatsa tsa lefutso tse phahameng tsa GC
4.Tatelano ea DNA ea boemo ba nanogram.
Boemo ba polokelo
Lipalangoang ka tlas'a 0°C 'me li bolokoe ho -25°C~-15°C.
Tlhaloso ea Yuniti
Yuniti e le 'ngoe e hlalosoa e le palo ea enzyme e kenyelletsang 25 nmol ea dNTP linthong tse sa keneng tsa asiti ka metsotso e 30 ho 65 ° C.
Taolo ea Boleng
1.Protein Purity Assay (SDS-PAGE):Bohloeki ba Bst DNA polymerase V2 ke ≥99% e khethiloeng ke tlhahlobo ea SDS-PAGE ho sebelisoa Coomassie Blue discovery.
2.Eke nyutlelieKetsahalo:Ho kenyeletswa ha 50 μL karabelo e nang le bonyane ba 8 U of Bst DNA polymerase V2 e nang le 1 μg λDNA bakeng sa dihora tse 16 ka 37 ℃ e fella ka hore ho se be le tshenyeho e lemohuwang jwalo ka ha ho laetswe.
3.Ts'ebetso ea Exonuclease:Ho kenyeletswa ha 50 μL reaction e nang le bonyane ba 8 U of Bst DNA polymerase V2 e nang le 1 μg λ -Hind Ⅲ digest DNA bakeng sa dihora tse 16 ka 37 ℃ e fella ka hore ho se be le tshenyeho e lemohuwang jwalo ka ha ho laetswe.
4.Mosebetsi oa Nickase:Ho kenyeletswa ha 50 μL karabelo e nang le bonyane ba 8 U of Bst DNA polymerase V2 e nang le 1 μg pBR322 DNA bakeng sa dihora tse 16 ka 37°C e fella ka hore ho se be le tshenyeho e lemohuwang jwalo ka ha ho hlalositswe.
5.Mosebetsi oa RNase:Ho kenyeletswa ha 50 μL karabelo e nang le bonyane ba 8 U of Bst DNA polymerase V2 e nang le 1.6 μg MS2 RNA bakeng sa dihora tse 16 ka 37°C e fella ka hore ho se be le tshenyeho e lemohuwang jwalo ka ha ho hlalositswe.
6.E. coliDNA:120 U of Bst DNA polymerase V2 e hlahlojoa boteng ba E. coli genomic DNA e sebelisa TaqMan qPCR e nang le li-primers tse tobileng bakeng sa sebaka sa E. coli 16S rRNA.Tšilafalo ea E. coli genomic DNA ke ≤1 Kopi.
LEMPA tsiboso
| Likaroloana | 25μL |
| 10 × HC Bst V2 Buffer | 2.5 μL |
| MgSO4 (100mM) | 1.5 μL |
| dNTPs (10mM e 'ngoe le e 'ngoe) | 3.5 μL |
| SYTO™ 16 e tala (25×)a | 1.0 μL |
| Motsoako oa peleb | 6 μL |
| Bst DNA Polymerase V2 (Glycerol-mahala) (8 U/uL) | 1 μL |
| Setšoantšo | × μL |
| ddH₂O | Ho fihlela ho 25 μL |
Lintlha:
1) a.SYTOTM 16 Green (25 ×): Ho ea ka litlhoko tsa liteko, li-dyes tse ling li ka sebelisoa e le li-substitutes;
2) b.Primer mix: e fumanoe ka ho kopanya 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 le mefuta e meng.
Boitšoaro le Boemo
1 × HC Bst V2 Buffer, mocheso oa incubation o pakeng tsa 60°C le 65°C.
Ho se sebetse ha mocheso
80°C, metsotso e 20
