Robustart Taq DNA Polymerase
Robustart Taq DNA Polymerase inopisa kutanga DNA polymerase.Ichi chigadzirwa hachingogone kuvharidzira zvisiri-chaiyo maitiro anokonzerwa nekusanyatso jekeswa kweprimers kana primer aggregation mukuita kwePCR system kugadzirira uye kukwidziridzwa.Naizvozvo, ine yakanakisa hunhu uye inoshanda zvakanyanya mukukwidziridzwa kweyakaderera matemplate matemplate, uye inokodzera multiplexed PCR amplification reaction.Zvakare, ichi chigadzirwa chine yakanaka kwazvo kushanda, uye yakagadzikana mibairo yekusimudza inogona kuwanikwa mumhando dzakasiyana dzePCR maitiro.
Zvikamu
1.5 U/μL Robustart Taq DNA polymerase
2.10 × PCR Buffer II (Mg²+ yemahara) (sarudzo)
3.25 mM MgCl2(sarudzo)
* 10 × PCR Buffer II (Mg²+ yemahara) haina dNTP neMg²+, ndapota wedzera dNTPs uye MgCl2paunenge uchigadzirira reaction system.
Recommended Applications
1.Kukurumidza kuwedzera.
2.Multiple amplification.
3.Kuwedzeredza kwakananga kweropa, swabs, uye mamwe masampuli.
4.Kuonekwa kwezvirwere zvekufema.
Storage Condition
-20°C kuitira kuchengetedza kwenguva refu, inofanira kusanganiswa zvakanaka isati yashandiswa, dzivisa kugaro chando-nyorova.
*Kana kunaya kwemvura kukaitika mushure mefiriji, zvakajairika;inokurudzirwa kuenzana nekamuri tembiricha usati wasanganisa uye kushandisa.
Chikamu Tsanangudzo
One active unit (U) inotsanangurwa sehuwandu hwe enzyme inobatanidza gumi nmol ye deoxyribonucleotide mu acid-insoluble material pa 74°C for 30mins using activated salmon urume DNA se template/primer.
Quality Control
1.SDS-PAGE electrophoretic kuchena kukuru kupfuura 98%.
2.Amplification senitivity, batch-to-batch control, kugadzikana.
3.Hapana exogenous nuclease chiitiko, hapana exogenous endonuclease kana exonuclease kusvibiswa.
Mirayiridzo
Reaction Setup
| Zvikamu | Vhoriyamu (μL) | Final Concentration |
| 10 × PCR Buffer II (Mg²+ yemahara)a | 5 | 1× |
| dNTPs (10mM imwe neimwe dNTP) | 1 | 200 μM |
| 25 mM MgCl2 | 2-8 | 1-4 mM |
| Robustart Taq DNA Polymerase (5U/μL) | 0.25-0.5 | 1.25-2.5 U |
| 25 × Primer musanganiswab | 2 | 1× |
| Template | - | ＜ 1 μg/kuita |
| ddH2O | kusvika ku50 | - |
Notes:
1) a.Iyo buffer haina dNTP neMg²+, ndapota wedzera dNTPs uye MgCl2paunenge uchigadzirira reaction system.
2) b.Kana yakashandiswa qPCR/qRT-PCR, fluorescent probes inofanira kuwedzerwa kune reaction system.Kazhinji, yekupedzisira primer concentration ye 0.2 μM ichapa mhedzisiro yakanaka;kana maitiro ekuita asina kunaka, iyo primer concentration inogona kugadziriswa muhuwandu hwe 0.2-1 μM.Iyo probe concentration inowanzogadziriswa muhuwandu hwe 0.1-0.3 μM.Concentration gradient miedzo inogona kuitwa kuti uwane yakanakisa musanganiswa weprimer uye probe.
Thermal cycling protocol
| Nguva dzose PCRprocess | |||
| Danho | Tembiricha | Nguva | Cycles |
| Pre-denaturation | 95℃ | 1-5 mins | 1 |
| Denaturation | 95℃ | 10-20 sec | 40-50 |
| Kuwedzera / Kuwedzera | 56-64 ℃ | 20-60 sec | |
| Fast PCRprocess | |||
| Danho | Tembiricha | Nguva | Cycles |
| Pre-denaturation | 95℃ | 30 sec | 1 |
| Denaturation | 95℃ | 1-5 sec | 40-45 |
| Kuwedzera / Kuwedzera | 56-64 ℃ | 5-20 sec | |
Notes
1.Mwero wekusimudzira wekukurumidza DNA polymerase haifanire kunge iri pasi pe1 kb/10 s.Iyo tembiricha yekukwira uye kudonha mwero, tembiricha yekudzora modhi uye kupisa conduction kushanda kweakasiyana PCR zviridzwa zvinosiyana zvakanyanya, saka zvinokurudzirwa kukwidziridza iyo yakakwana yekuita mamiriro eiyo chaiyo inokurumidza PCR chiridzwa.
2.Iyo sisitimu inochinjika zvakanyanya, ine hunyanzvi hwepamusoro uye kunzwa.
3.Inokodzera kushandiswa seyakanyanya senitivity PCR yekuona reagents, uye inogona kushandiswa mumultiplex PCR amplification maitiro.
4.5′→3′ polymerase chiitiko, 5′→3′ exonuclease chiitiko;kwete 3′→5′ exonuclease basa;hapana basa rekuongorora.
5.Inokodzera kuongororwa kwemhando uye kuwanda kwePCR uye RT-PCR.
6.Iyo 3 ′ yekupedzisira yePCR chigadzirwa ndeye A, iyo inogona kuumbwa yakananga muT vector.
7.Iyo nhanho-nhatu nzira inokurudzirwa kune maprimers ane yakaderera annealing tembiricha kana yekukudza zvidimbu zvinodarika 200 bp.
