Animal Tissue Direct PCR kit (ntle le Sampling Tools) Protocol
Tlhaloso ea Kit:
Sebelisa lysate ea linama tsa liphoofolo ka kotloloho joalo ka template ea ho holisa PCR, ntle le ho ntšoa ka thoko ho DNA, kutlo e potlakileng le e phahameng.
◮Ha ho hlokahale tlhoekiso ea DNA e jang nako le e theko e boima.
◮Tlhokahalo ea sampole e nyane, nka peo e le 'ngoe feela.
◮Ha ho hlokahale liphekolo tse khethehileng tse kang ho sila le ho sila, 'me ho buuoa ho bonolo.
◮Sistimi e ntlafalitsoeng ea PCR e thusa PCR ho ba le maemo a holimo le mamello e matla ho PCR reaction inhibitors.
Litlhaloso
Setsi sena se sebelisa sistimi e ikhethileng ea lysis buffer ho lokolla ka potlako DNA ea lisele tsa liphoofolo bakeng sa karabelo ea PCR, ka hona e loketse haholo tlhahlobo e kholo ea liphatsa tsa lefutso.
Mokhoa oa ho lokolla DNA ea genomic ho tswa ho lysis buffer e phethoa ka hare ho metsotso e 10-30 ho 65 ° C.Ha ho na mekhoa e meng e kang ho tlosoa ha protheine le RNA e hlokahalang, 'me DNA ea trace e lokolotsoeng e ka sebelisoa e le template bakeng sa karabelo ea PCR.
2 × PCR e bonoloTMMix e na le mamello e matla ho PCR reaction inhibitors, 'me e ka sebelisa lysate ea sampole ho lekoa e le template bakeng sa ho holisa ka nepo le ka nepo.Reagent ena e na le Foregene D-Taq DNAPolymerase, dNTPs, MgCl.2, buffer ea karabelo, PCR optimizer le stabilizer.
D-Taq DNA polymerase ke DNA polymerase e entsoeng ka ho khetheha ke Foregene bakeng sa karabelo e tobileng ea PCR.D-Taq DNA polymerase e na le mamello e matla ho mefuta e fapaneng ea li-inhibitors tsa PCR, 'me e khona ho holisa DNA ka mokhoa o nepahetseng lits'ebetsong tse fapaneng tse rarahaneng tsa karabelo, mme lebelo la amplification le ka fihla ho 2Kb/min, e loketseng ka ho khetheha bakeng sa karabelo ea Direct PCR.
Litlhaloso
50×20µl rxns, 200×20µl rxns, 500×20µl rxns, 2000×20µl rxns
Likarolo tsa Kit
| Karolo ea I | Buffer AL |
| Foregene Protease | |
| 6× DNA Loading Buffer | |
| Karolo ea II | 2 × PCR e bonoloTMKopanya |
Likarolo le melemo
■Ha ho tlhoekiso ea DNA e jang nako le e theko e boima
■ Tlhokahalo ea sampole e nyane, joalo ka ha 5mg lesela la liphoofolo le ka lekoa.
■ Ha ho na phekolo e khethehileng e kang ho sila le ho sila e hlokahalang, 'me ho buuoa ho bonolo.
■ Sistimi e ntlafalitsoeng ea PCR e etsa hore PCR e be le maemo a holimo le mamello e matla ho PCR reaction inhibitors.
Litekanyetso tsa kit
Application: mefuta e fapaneng ya dinama tse nyenyane tse phoofolo.
DNA e lokollotsoeng ho tsoa ho sampole lysis: e sebelisoa feela joalo ka template ea PCR.
Thepa e ka sebelisoa bakeng sa merero e latelang: ho tsebahatsa transgenosis, genotyping ea liphoofolo, joalo-joalo.
Phallo ea mosebetsi
Storage le Shelf bophelo
Karolo ea I ea kit ena e lokela ho bolokoa ho 2-8℃.
Reagent Buffer AL e ka bolokoa likhoeli tse 12 tlas'a maemo a omileng;e ka bolokoa ho 2-8℃bakeng sa polokelo e telele.
Reagent Foregene Protease e na le foromo e ikhethang.Ho netefatsa ts'ebetso ea eona le botsitso, ka kopo e boloke ho 4℃bakeng sa likhoeli tse 12.
Reagent 6×DNA Loading Buffer e ka bolokoa ho 4℃kapa -20℃bakeng sa nako e telele.
Karolo ea II ea kit ena e lokela ho bolokoa ho -20℃.
Reagent 2×PCR e bonoloTMMotsoako o ka bolokoa likhoeli tse 12 ho -20℃;haeba e sebelisoa khafetsa, e ka bolokoa ho 4℃bakeng sa polokelo ea nako e khuts'oane (sebelisa ho fihlela matsatsing a 10).
