Inyama zinyamanswa Direct PCR kit-UNG (idafite ibikoresho byo gutoranya) Porotokole
Ibisobanuro:
Koresha mu buryo butaziguye inyamanswa yinyamanswa nkicyitegererezo cyo kongera PCR, udakuyemo ADN zitandukanye, byihuse kandi byoroshye.Wongeyeho UNG yo kurwanya umwanda kugirango ikureho ibyiza.
◮Ntabwo bitwara igihe kandi bihenze bya ADN bisabwa.
◮Icyitegererezo gisabwa ni gito, fata imbuto imwe.
◮Nta buvuzi budasanzwe nko gusya no kumenagura bisabwa, kandi imikorere iroroshye.
◮Sisitemu nziza ya PCR ituma PCR igira umwihariko wo kwihanganira no kwihanganira imbaraga za PCR.
Ibisobanuro
Iki gikoresho gikoresha sisitemu yihariye ya lysis kugirango irekure vuba ADN ya genomic kuva mubitangangingo byinyamanswa kubitekerezo bya PCR, kubwibyo birakwiriye cyane cyane kwipimisha rinini.
Igikorwa cyo kurekura ADN genomic muri buffer ya lysis kirangiye muminota 10-30 kuri 65 ° C.Ntayindi nzira nka protein na RNA ikuraho, kandi ADN yarekuwe irashobora gukoreshwa nkicyitegererezo cya PCR.
2 × PCR BiroroshyeTMKuvanga (UNG) ikoresha dUTP aho gukoresha dTTP hashingiwe kuri 2 × PCR ByoroshyeTMKuvanga, hanyuma ukongeramo enzyme ya UNG (Uracil-N-glycosylase) ishobora gutesha agaciro inyandikorugero irimo dUTP icyarimwe.Mbere ya PCR reaction, enzyme ya UNG ikoreshwa mugutesha agaciro ibicuruzwa bya PCR birimo uracil.Enzyme ya UNG ntacyo izagira ku gishushanyo kitarimo uracil, bityo hakemezwa umwihariko nukuri kwokwiyongera no gukumira ko ibicuruzwa bya PCR byandura mugihe kinini cyo gupima geneti.
D-Taq ADN polymerase ni ADN polymerase yakozwe na Foregene kugirango reaction ya PCR itaziguye.D-Taq ADN polymerase yihanganira cyane ibintu bitandukanye byangiza PCR, kandi irashobora kongera imbaraga za ADN muri sisitemu zitandukanye zitoroshye, kandi umuvuduko wa amplification ushobora kugera kuri 2Kb / min, cyane cyane muburyo bwa PCR.
Ibigize ibikoresho
| Igice cya I. | Buffer AL |
| Foregene Protease | |
| 6 × ADN Yipakurura Buffer | |
| Igice cya II | 2 × PCR BiroroshyeTMKuvanga (UNG) |
Ibiranga ibyiza
■Ntabwo bitwara igihe kandi bihenze byo kweza ADN
Require Icyitegererezo gisabwa ni gito, nkibice 5mg byinyamanswa bishobora kugeragezwa.
■ Nta buvuzi budasanzwe nko gusya no kumenagura bisabwa, kandi imikorere iroroshye.
System Sisitemu nziza ya PCR ituma PCR igira umwihariko wo kwihanganira no kwihanganira imbaraga za PCR.
System Sisitemu yo kurwanya umwanda PCR 2×PCR BiroroshyeTMKuvanga (UNG) bikuraho neza umwanda uterwa nibicuruzwa bya PCR kandi ukemeza umwihariko nukuri kwa amplification.
Ibipimo by'ibikoresho
Gushyira mu bikorwa: inyama zinyamaswa zitandukanye.
ADN yasohotse muri sample lysis: ikoreshwa gusa nkicyitegererezo cya PCR.
Igikoresho kirashobora gukoreshwa mubikorwa bikurikira: kumenya transgenose, genotyping yinyamaswa, nibindi.
Urujya n'uruza rw'akazi
Ububiko hamwe nubuzima bwa Shelf
Igice cya I cyiki gitabo kigomba kubikwa kuri 2-8℃.
Buffer AL reagent irashobora kubikwa amezi 12 mugihe cyumye;irashobora kubikwa kuri 2-8℃kubikwa igihe kirekire.
Reagent Foregene Protease ifite formula idasanzwe.Kugirango ibikorwa byayo bihamye, nyamuneka ubibike kuri 4℃amezi 12.
Reagent 6×ADN Yipakurura Buffer irashobora kubikwa kuri 4℃cyangwa -20℃igihe kirekire.
Igice cya II cyiki gikoresho kigomba kubikwa kuri -20℃.
Reagent 2×PCR BiroroshyeTMKuvanga (UNG), niba ikoreshwa kenshi, irashobora kandi kubikwa kuri 4℃kubikwa igihe gito (koresha muminsi 10).
