Mouse Tail Direct PCR Kit-UNG Direct PCR Lysis Reagent(Mouse Tail)(bakeng sa Genotyping)
Litlhaloso
Setsi sena se sebelisa mokhoa o ikhethileng oa lysis buffer system ho lokolla DNA ea genomic ka potlako mehatleng ea litoeba, litsebe, mesifa le disampole tse ling tsa lisele 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 × M1-PCR e BonoloTMMix (UNG) e sebelisa dUTP sebakeng sa dTTP motheong oa 2× M1-PCR EasyTM Kopanya (UNG), mme o eketsa UNG enzyme (Uracil-N-glycosylase) e ka senyang template e nang le dUTP ka nako e le 'ngoe.Pele ho karabelo ea PCR, enzyme ea UNG e sebelisoa ho nyenyefatsa sehlahisoa sa PCR se nang le uracil.Enzyme ea UNG e ke ke ea e-ba le phello leha e le efe ho template e se nang uracil, ka tsela eo e netefatsa ho nepahala le ho nepahala ha amplification le ho thibela monyetla oa tlhahlobo e kholo ea liphatsa tsa lefutso.Tšoaetso ea lihlahisoa tsa PCR e etsahetse.
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 MP |
Foregene Protease Plus | |
6× DNA Loading Buffer | |
Karolo ea II | 2× M1 PCR e BonoloTMMotsoako(UNG) |
Litaelo |
Likarolo le melemo
■Ha ho hlokahale tlhoekiso ea DNA e jang nako le e theko e boima.
■ Tlhokahalo ea sampole e nyane, hanyenyane joalo ka 5mg toeba kapa mohatla oa 2-5mm o ka lekoa.
■ Ha ho hlokahale liphekolo tse khethehileng tse kang ho sila le ho sila, ’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.
■ Sistimi ea PCR e thibelang tšilafalo 2×M1-PCR e bonoloTMMix (UNG), e felisang ka katleho tšilafalo e bakoang ke lihlahisoa tsa PCR le ho netefatsa ho nepahala le ho nepahala ha amplification.
Litekanyetso tsa kit
Sebaka sa ts'ebeliso: mohatla oa mouse, tsebe ea mouse, mesifa le lisele tse ling.
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℃ nako e telele.
Karolo ea II ea kit ena e lokela ho bolokoa ho -20 ℃.
Reagent 2 × M1 PCR EasyTM Mix(UNG), haeba e sebelisoa khafetsa, e ka boela ea bolokoa ho 4 ℃ bakeng sa polokelo ea nako e khutšoanyane (sebelisa pele ho matsatsi a 10).