Ungakhetha kanjani izinto ezisetshenziswayo ze-PCR/qPCR?

I-PCR iyindlela ebucayi nesebenzayo yokukhulisa ikhophi eyodwa yokulandelana kwe-DNA eqondiwe ezigidini zamakhophi ngesikhathi esifushane. Ngakho-ke, izinto ezisetshenziswayo zepulasitiki zokusabela kwe-PCR kumele zingabi namagciwane nama-inhibitor, kuyilapho zibe nekhwalithi ephezulu engaqinisekisa umphumela ongcono kakhulu we-PCR. Izinto ezisetshenziswayo zepulasitiki ze-PCR zitholakala ngamasayizi namafomethi ahlukahlukene, futhi ukwazi izici ezifanele zemikhiqizo kuzokusiza ekukhetheni izinto ezifanele zepulasitiki ezisebenzisekayo ukuze uthole idatha ye-PCR ne-qPCR efanele.

Ukwakheka kanye Nezimpawu ze-PCR ezisebenzisekayo

1.Izinto
Izinto ezisetshenziswayo ze-PCR ngokuvamile zenziwe nge-polypropylene, engasebenzi ngokwanele ukuze imelane nokushintsha kwezinga lokushisa ngokushesha phakathi nokuhamba ngebhayisikili eshisayo futhi kuncishiswe ukutholwa kwezinto ezisebenzayo ukuze kuqinisekiswe imiphumela emihle ye-PCR. i-medical-grade, i-polypropylene yekhwalithi ephezulu kufanele isetshenziswe ngesikhathi sokukhiqiza futhi yenziwe endaweni yokuhlanza ye-Class 100,000. Umkhiqizo kufanele ungabi namagciwane kanye ne-DNA ukuze ugweme ukuphazamisa umphumela wokuhlolwa kokukhulisa i-DNA.

2.Umbala
Amapuleti e-PCRfuthiAmashubhu e-PCRngokuvamile atholakala ngokusobala nokumhlophe.

• Idizayini yokuqina kodonga olufanayo izohlinzeka ngokudluliswa kokushisa okungaguquki kwamasampuli asabelayo.

• Ukungena okuphezulu kwe-optical ukuqinisekisa ukudluliswa kwesignali ye-fluorescence kanye nokuhlanekezela okuncane.

• Ocwaningweni lwe-qPCR, imbobo emhlophe ivimbele ukuphindwa kwesignali ye-fluorescence kanye nokumuncwa kwayo ngemojuli yokushisisa.

3.Ukufometha
I-PCR Plate "isiketi" sizungeze ibhodi. Isiketi sinikeza ukuzinza okungcono kwenqubo yepayipi lapho uhlelo lokusabela lwakhiwe, futhi lunikeze amandla angcono wemishini ngesikhathi sokwelashwa kwemishini okuzenzakalelayo. Ipuleti le-PCR lingahlukaniswa libe akukho isiketi, isiketi esiyingxenye nesiketi esigcwele.

• Ipuleti le-PCR elingagqokile alikho epuletini, futhi leli fomu lepuleti lokusabela lingashintshwa kumathuluzi amaningi e-PCR namamojula wethuluzi le-PCR lesikhathi sangempela, kodwa hhayi izinhlelo zokusebenza ezizenzakalelayo.

• Ipuleti le-PCR eline-semi-skirted linomkhawulo omfushane onqenqemeni lwepuleti, linikeza usekelo olwanele phakathi ne-pipetting namandla okusebenza okuphatha irobhothi.

• Ipuleti le-PCR elinesiketi esigcwele linomphetho ovala ubude bepuleti. Leli fomu lepuleti lilungele ukusebenza okuzenzakalelayo, okungaba ukuzivumelanisa okuphephile nokuzinzile. Isiketi esigcwele siphinde sithuthukise amandla emishini, siyenze ilungele ukusetshenziswa namarobhothi ekuhambeni komsebenzi okuzenzakalelayo.

Ishubhu ye-PCR itholakala ngeshubhu eyodwa kanye ne-8-strips, afaneleka kakhulu ekuhlolweni kokuphuma okuphansi kuya kokumaphakathi kwe-PCR/qPCR. Ikhava eyisicaba iklanyelwe ukwenza lula ukubhala, futhi ukudluliswa kokwethembeka okuphezulu kwesiginali ye-fluorescence kungenziwa kangcono nge-qPCR.

• Ishubhu elilodwa linikeza ukuguquguquka ukuze usethe inani eliqondile lokusabela. Kumavolumu amakhulu okusabela, ishubhu elilodwa likasayizi ongu-0.5 mL liyatholakala.
• Ishubhu elinemichilo engu-8 elinamakepisi livula futhi livale amashubhu esampula ngokuzimela ukuze kuvinjelwe isampula.

4.ukuvalwa
Isembozo seshubhu kanye nefilimu yokuvala kufanele ivale ngokuphelele ishubhu nepuleti ukuvimbela ukuhwamuka kwesampula phakathi nomjikelezo wokushisa. Isigxivizo esiqinile singabonakala ngokusebenzisa i-scraper yefilimu kanye nethuluzi lokucindezela.

• Imithombo yamapuleti e-PCR inomphetho ophakanyisiwe nxazonke. Lo mklamo usiza ukuvala ipuleti ngefilimu yokuvala ukuvimbela ukuhwamuka.
• Izimpawu ze-alphanumeric epuletini le-PCR zizosiza ukukhomba imithombo ngayinye kanye nokuma kwamasampula ahambisanayo. Izinhlamvu ezinamaqhubu ngokuvamile zinyatheliswa ngombala omhlophe noma omnyama, futhi ezinsizeni ezizenzakalelayo, ukubhala kunenzuzo enkulu ekuvaleni imiphetho yangaphandle yepuleti.

5.Flux isicelo

Ukuguquguquka kokuhlolwa kokuhlolwa kwe-PCR/qPCR kungase kunqume ukuthi yiluphi uhlobo lwezinto ezisetshenziswayo zepulasitiki okufanele zisetshenziselwe umphumela wokwelapha ongcono kakhulu. Ngohlelo lokusebenza lokuphuma okuphansi ukuya kokumaphakathi, amashubhu ngokuvamile afaneleka kakhulu, kuyilapho amapuleti edingeka kakhulu ekuhlolweni kokuphuma okuphakathi kuya kokuphezulu. Amapuleti aphinde aklanyelwe ukucabangela ukuguquguquka kokuguquguquka, okungahlukaniswa kube umugqa owodwa.

Sengiphetha, njengengxenye ebalulekile yokwakhiwa kwesistimu ye-PCR, izinto ezisebenzisekayo zepulasitiki zibalulekile empumelelweni yokuhlolwa nokuqoqwa kwedatha, ikakhulukazi ezinhlelweni zokusebenza zokugeleza komsebenzi okuphakathi kuya kokuphezulu.

Njengomphakeli waseShayina wezinto ezisetshenziswayo zepulasitiki ezizenzakalelayo, i-Cotaus inikeza amathiphu e-pipette, i-nucleic acid, ukuhlaziya amaprotheni, isiko lamaseli, isampula yokugcina, ukuvala uphawu, i-chromatography, njll.

Chofoza isihloko somkhiqizo ukuze ubuke imininingwane yomkhiqizo we-PCR.

PCR Tube ;Ipuleti le-PCR