2 × Rapid Taq Super Mix
Katsi Nhamba: HCR2016A
2 × Rapid Taq Super Musanganiswa yakavakirwa pane yakagadziridzwa Taq DNA Polymerase, ichiwedzera yakasimba yekuwedzera chinhu, amplification yekusimudzira chinhu uye yakagadziridzwa buffer system, ine yakanyanya kukwidziridza kunyatsoita.Iko kumhanya kwekukwidziridzwa kwematemplate akaoma senge genome mukati me3 kb inosvika 1-3 sec/kb, uye iyo yematemplate akareruka semaplasmids mukati me5 kb inosvika 1 sec/kb.Ichi chigadzirwa chinogona kuchengetedza zvakanyanya PCR yekuita nguva.Panguva imwecheteyo, musanganiswa une dNTP neMg2+, iyo inogona kukwidziridzwa chete nekuwedzera maprimers uye matemplate, ayo zvakare anorerutsa zvakanyanya matanho ekushanda ekuedza.Uyezve, musanganiswa une electrophoretic chiratidzo dhayi, inogona kuve yakananga electrophoresis mushure mekuita.Mushandi anodzivirira muchigadzirwa ichi anoita kuti musanganiswa uchengetedze basa rakagadzikana mushure mekudzokororwa kwechando uye kunyunguduka.Iyo 3'-yekupedzisira bhendi A yePCR chigadzirwa inogona kugadzirwa nyore muT vector.
Zvikamu
2 × Rapid Taq Super Mix
Storage Conditions
PCR Master Mix zvigadzirwa zvinofanirwa kuchengetwa pa -25~-15 ℃ kwemakore maviri.
Zvinotsanangurwa
| Product specification | Rapid Taq Super Mix |
| Concentration | 2× |
| Sravana Sameeralu Serial 4th Hot Start | Yakavakwa-mukati Hot Start |
| Overhang | 3′-A |
| Reaction speed | Rapid |
| Saizi (Chigadzirwa Chekupedzisira) | Kusvika ku15 kb |
| Mamiriro ekufambisa | Dry ice |
Mirayiridzo
1. Rection System (50 μL)
| Zvikamu | Saizi (μL) |
| DNA template* | zvakakodzera |
| Pamberi pekutanga (10 μmol/L) | 2.5 |
| Reverse primer (10 μmol/L) | 2.5 |
| 2 × Rapid Taq Super Mix | 25 |
| ddH2O | kusvika ku50 |
2.Amplification Protocol
| Matanho ekutenderera | Tembiricha (°C) | Nguva | Cycles |
| Predenaturation | 94 | 3 min | 1 |
| Denaturation | 94 | 10 sec |
28-35 |
| Annealing | 60 | 20 sec | |
| Extension | 72 | 1-10 sec/kb |
Inokurudzirwa kushandiswa kwematemplate akasiyana:
| Rudzi rwe template | Chikamu chekushandisa renji (50 μL reaction system) |
| Genomic DNA kana E. coli mvura | 10–1,000 ng |
| Plasmid kana viral DNA | 0.5-50 ng |
| cDNA | 1-5 µL (kwete kupfuura 1/10 yehuwandu hwehuwandu hwePCR reaction) |
| Inokurudzirwa kushandiswa kwematemplate akasiyana | |
Notes:
1.Kushandisa Reagent: nyungudika zvizere uye sanganisa usati washandisa.
2. Annealing tembiricha: The annealing tembiricha ndiyo yepasirese Tm kukosha, uye inogona zvakare kusetwa 1-2 ℃ yakaderera pane primer Tm kukosha.
3. Kuwedzera kukurumidza: Isa 1 sec/kb yematemplate akaoma akadai segenome uye E. coli mukati me1 kb;set 3 sec/kb for complex templates se 1-3 kb genome uye E. coli;set 10 sec/kb for complex templates pamusoro pe3 kb genome neE. coli.Unogona kuseta kukosha ku1 sec/kb kune template iri nyore senge plasmid isingasviki 5 kb, 5 sec/kb kune iri nyore template senge plasmid iri pakati pe5 ne10 kb, uye 10 sec/kb ye template iri nyore. senge plasmid yakakura kupfuura 10 kb.
Notes
1. Nekuchengetedza uye hutano hwako, ndapota pfeka majasi erabhu uye magirovhosi anoraswa kuti ushande.
2. Ichi chigadzirwa ndechekutsvaga kushandiswa CHETE!
