Senza futhi ucwaningo lwe-PCR, kungani imiphumela ihlale imbi kakhulu kuneminye?

Senza futhi ucwaningo lwe-PCR, kungani imiphumela ihlale imbi kakhulu kuneminye?

Elabhorethri, bheka i-PCR yabanye abantu, ukusebenza kwe-amplification kuphezulu, umucu we-electrophoresis uphelele, bese ubheka okwabo, okunzima ukukwenza isikhathi eside, uzwe ukuthi ukusabela kuhle kakhulu, umphumela wesivivinyo esingelona iqiniso …… Yini inkinga?

Ngikholwa wukuthi iningi lozakwethu nalo lihlangabezane nezinkinga ezahlukahlukene. Lapho singena odongeni ekuhlolweni kwe-PCR, sizohlaziya ngokucophelela izinto ezingaphakathi kohlelo lokuphendula, kepha sivame ukungalinaki ithonya elibalulekile lokuthengiswayo ekuhlolweni.

Umphumela wakho we-PCR awulungile, okudlekayo kwe-PCR kungaba yisizathu esibalulekile. Ngakolunye uhlangothi, ukungcoliswa okuncane kwezinto ezisetshenziswayo noma ukwethulwa kwama-inhibitors kuzodala ukungcoliswa kokuhlola; Ngakolunye uhlangothi, ukukhethwa okungafanele kokuthengiswayo kuzokwenza imiphumela yokuhlola ihlupheke.

Inkinga iza futhi. Kunezinhlobo eziningi ze-PCR ezisetshenziswayo, ngakho-ke ungazikhetha kanjani ezifanele ukwenza ukuhlolwa kube bushelelezi? Yiziphi izinkinga okufanele zinakwe? Ungakhathazeki, namuhlaI kuhlanganiswe neminyaka eminingi yokuhlangenwe nakho, ukuhlunga izinkinga ezikhethiwe zokusebenzisa i-PCR, ngithemba ukukusiza.

 

I-Q1: Kungani ukusetshenziswa kwe-PCR kuvame ukwenziwa ngezinto ze-PP?

I-Huida: Ngoba izimpahla ze-PCR zivame ukuthintana ngqo nama-reagents noma amasampuli, izinto ze-polypropylene (PP) azinazinto eziphilayo, azilandeli ama-biomoleculars, futhi zinokubekezelelana okuhle kwamakhemikhali nokubekezelelana kwezinga lokushisa (kungavalwa ngokuzenzakalela ngo-121 ° C, futhi ingamelana nezinguquko zokushisa phakathi nomjikelezo oshisayo).

 

I-Q2: Kufanele ngikhethe kanjani phakathi kosayizi abahlukahlukene bamashubhu / amapuleti e-PCR?

I-HUIDA: Inhloso yokukhetha: khetha imikhiqizo efanele ngokuya ngezidingo ezithile zokuhlola. Iningi losayizi webhubhu le-PCR lingahlangabezana nezidingo zokuphendula kwe-PCR. Ngalesi sizathu, kunconywa ukuthi ukhethe ithubhu yevolumu ephansi kuqala. Ngoba i-tube / ipuleti yokuphendula yevolumu ephansi inesikhala esincane ngaphezulu, ukuqhutshwa kokushisa kuyathuthukiswa futhi ukuhwamuka kuncishisiwe. Ngasikhathi sinye, kuyadingeka ukugwema ukufaka amasampula amaningi kakhulu noma amancane kakhulu. Ukuningi kakhulu kungaholela ekwehlisweni kokushisa okushisayo, ukuchitheka nokungcola okuphambene, ngenkathi kuncane kakhulu kungaholela ekulahlekelweni kwesampula kokuhwamuka.

Imininingwane kanye nenani lamashubhu wokuphendula ajwayelekile:

Ishubhu elilodwa / elihlanganisiwe: 0.5ml, 0.2ml, 0.15ml

I-96-well plate: 0.2ml, 0.15ml

Ipuleti elinemithombo engu-384: 0.04ml

 

I-Q3: Lapho inani lesampula lincane, ngizokhetha ipayipi elilodwa noma ipayipi elihlanganisiwe, kepha kungani amanye enekhava eyisicaba kanti amanye anekhava ye-convex?

I-Huida: Lapho inani lesampula lincane, ithubhu elilodwa noma ishubhu elihlanganisiwe lizokhethwa. Kodwa-ke, lapho ivolumu yokuphendula inkulu, ishubhu elilodwa libusa, futhi ivolumu ingafinyelela ku-0.5 mL. Futhi ikhava eyisicaba nesembozo esihlanganayo ngasinye sinenzuzo, okuqondile kungabhekisa kulwazi olulandelayo.

Ishubhu elilodwa: 0.2ml no-0.5ml; Ungakhetha ubuningi besampula kalula.

Ukuxhuma ishubhu: 0.2ml noma u-0.15ml kungakhethwa; Amashubhu ayi-8 noma ayi-12 ajwayelekile.

Ikhava Flat: anganikeza ukudluliswa kwesiginali se-fluorescence esinembile se-qPCR; Kulula ukubhala amamaki.

Ikhava ye-Convex: ukuxhumana nesembozo esishisayo sethuluzi le-PCR ukunciphisa ukusikeka kwethumbu lokuphendula elibangelwa ingcindezi; Kodwa-ke, izothinta ukudluliswa kwesiginali ye-fluorescence futhi ayikwazi ukusetshenziswa ekuhlolweni kwe-qPCR.

 

I-Q4: Kungani amanye amapuleti e-PCR eneziketi kanti amanye awanazo?

I-Huida: Eqinisweni, isiketi sepuleti le-PCR senzelwe ukujwayela kangcono izinhlelo zokusebenza ezishintshayo, ukuhlinzeka ukwesekwa okuzinzile nokukhuthazela ngomshini kwensimbi, kanye nokuzinza okuphezulu ngesikhathi senqubo yokubhobhoza.

Amapuleti e-PCR ngokuvamile ahlukaniswe abe yi-non-hemline, half-hemline ne-full-hemline.

Ngaphandle kwe- Ipuleti: Ilungele imishini eminingi ye-PCR noma imishini ye-qPCR, kepha hhayi izinhlelo zokusebenza ezishintshayo. Ukuzinza akuphakeme kunqubo yokufaka amapayipi, ngakho-ke kudinga ukuthi kusetshenziswe ngokuxhaswa kwepuleti.

Ipuleti onqenqemeni lwesiketi: ingashintshwa ukuze ifake ilebula noma ikhodi yebha, nohlelo oluzenzakalelayo, futhi inokuzinza okuhle kwepayipi.

Ipuleti elinqenqemeni lesiketi eligcwele: lilungele kakhulu izinhlelo zokusebenza ezizenzakalelayo zokuhlola, nalo lingashintshwa liye kulebula nasekusebenziseni ikhodi yebha. Inamandla amahle wokukhanda, ingasetshenziswa ku-module ye-bulge module ye-PCR, futhi inokuzinza okuphezulu kunqubo yokupayipha.

 

Q5 :Kumapuleti e-PCR, kungani amakhona namalebula kungafani ncamashi?

I-Huida: Kuya ngomphumela wokusika kwekona nokumaka.

Ukuzisika kwekhoneni: Ukukhethwa kwendawo yokusika ekhoneni lepuleti le-PCR kuncike kuzidingo zensimbi efanelekayo yokubeka kalula.

Ukuhlonza: Omaka be-Alphanumeric kuma-PCR plate basiza ukukhomba ama-Wells ngamanye nezindawo zesampula. Ngokuvamile embossed umbala logo digital noma logo libhalwe. Kwezinye izinhlelo zokusebenza ezizenzakalelayo, amapuleti okuphendula amakiwe amakwe kangcono.

 

I-Q6: Kungani amanye amapuleti e-PCR eyi-convex kanti amanye ayizicaba? Yikuphi okungcono?

U-Huida: Okulingana kungcono kakhulu. Kunezinhlobo ezimbili zamapuleti ajwayelekile e-PCR: i-flat pore margin ne-pore margin ephezulu.

Ipuleti lokuphendula lemajini elincane, elifanele iningi lamathuluzi we-PCR.

Amapuleti okuphendula anemiphetho ye-orifice ekhulayo enza kube lula ukubekwa uphawu kolwelwesi futhi kunciphise ubungozi bokungcola okuwela phakathi kwamasampuli.

 

U-Huida unolwazi lweminyaka eminingi ekukhiqizweni nasekucubungulweni kwemikhiqizo ye-PCR nezindinganiso eziphakeme zemboni. Ulayini womkhiqizo ucebile futhi ungahlangabezana neningi lezinhlelo zokusebenza zokuhlola ze-PCR. Woza ukhethe imikhiqizo efanele yezinhlelo zakho zokusebenza zokuhlola.

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Isikhathi Iposi: Jun-03-2021