Ukuxazulula ukuxazulula ukuxazulula izinkinga: imibuzo ebuzwa njalo nezixazululo

Abahlaziyi be-Polymerase Chain Reaction (PCR) bangamathuluzi abalulekile ku-Bilefanelar Biology, evumela abaphenyi ukuthi bakhulise i-DNA ukuthola izinhlelo zokusebenza ezisukela ekuhlolweni kwezakhi zofuzo. Kodwa-ke, njenganoma iyiphi insiza eyinkimbinkimbi, i-PCR analyzer ingahlangabezana nezinkinga ezithinta ukusebenza kwayo. Le ndatshana ibhekana nemibuzo evamile mayelanaUkuhlaziywa kwe-PCRUkuxazulula inkinga futhi kunikeza izixazululo ezisebenzayo ezinkingeni ezijwayelekile.

1. Kungani ukusabela kwami ​​kwe-PCR akukhulisi?

Enye yezinkinga ezivame kakhulu abasebenzisi ababhekene nabasebenzisi ukukwazi ukusabela kwe-PCR ukukhulisa i-DNA eqondiwe. Lokhu kungabikwa ngezinto eziningana:

Umklamo we-primer engalungile Sebenzisa amathuluzi wesoftware e-Primer Design ukugwema ukubopha okungenangqondo.

I-Pempulate yethempulethi enganele: qinisekisa ukuthi usebenzisa inani elanele le-template DNA. Kuncane kakhulu kuzophumela ekulweni okubuthakathaka noma akukho.

I-inhibitors kusampula: Ukungcola kusampula kungavimba ukusabela kwe-PCR. Cabanga ukuhlanza i-DNA yakho noma usebenzisa indlela ehlukile yokukhipha.

Isixazululo: Hlola ukwakheka kwakho kwe-Primer, ukhulise ukugxila kwethempulethi, futhi uqiniseke ukuthi isampula yakho ayiqukethe ama-inhibitors.

2. Kungani umkhiqizo wami we-PCR usayizi ongalungile?

Uma usayizi wakho womkhiqizo we-PCR ungalindelekile, kungakhombisa inkinga ngezimo zokuphendula noma izithako ezisetshenzisiwe.

Ukukhuliswa okungelona obalulekile: Lokhu kungenzeka uma i-primer ibopha kwisiza esingahlosiwe. Bheka ukucaciswa kwama-primers usebenzisa ithuluzi elifana nokuqhuma.

Izinga lokushisa elingalungile Ukusebenziseka kahle kwezinga lokushisa lokungena nge-gradient PCR.

Isixazululo: Qinisekisa ukucaciswa kwe-Primer futhi uthuthukise izinga lokushisa lokuvuselela ukuthuthukisa ukunemba kwemikhiqizo ye-PCR.

3. I-My PCR Analyzer ikhombisa umyalezo wephutha. kumele ngenzeni?

Imilayezo yephutha kwi-PCR Analyzer ingashaqisa, kepha ngokuvamile inganikeza izinkomba ezinkingeni ezingaba khona.

Izinkinga zokulinganisa: qiniseka ukuthi i-PCR Analyzer ilinganiselwe kahle. Ukulungiswa njalo kanye namasheke wokulinganisa kubalulekile ekutholeni imiphumela efanele.

Iqembu lesoftware: Kwesinye isikhathi, izimbungulu zesoftware zingadala izinkinga. Qala kabusha ikhompyutha yakho bese uhlola izibuyekezo zesoftware.

Isixazululo: Bheka incwajana yomsebenzisi yekhodi ethile yephutha bese ulandela izinyathelo ezinconyiwe zokuxazulula izincomo. Ukulungiswa okujwayelekile kungavimbela izinkinga eziningi.

4. Kungani imiphumela yami yokuphendula ye-PCR ingahambisani?

Imiphumela ye-PCR engahambelani ingakhungathekisa ngenxa yezizathu eziningana:

Ikhwalithi ye-REEGENT: Qinisekisa ukuthi yonke i-reagents, kufaka phakathi ama-enzyme, ama-buffers, nama-DNTPs, amasha futhi aphezulu. Ama-reagents aphelelwe yisikhathi noma angcolisiwe angadala ukuhlukahluka.

I-Thermal cyper calibration: Izilungiselelo zokushisa ezingahambisani zingathinta inqubo ye-PCR. Hlola njalo ukulinganisa kwe-cyper ye-thermal.

Isixazululo: Sebenzisa ama-reagents asezingeni eliphakeme futhi ulinganise i-cycler yakho ye-thermal njalo ukuze uqinisekise imiphumela engaguquki.

I-5. Ungakuthuthukisa Kanjani Ukusebenza Kokuphendula kwe-PCR?

Ukwenza ngcono ukusebenza kahle kwezindlela ze-PCR kungaholela ekuvutheni okuphezulu nemiphumela ethembekile.

Lungiselela izimo zokuphendula: Ukuhlola usebenzisa ukugxila okuhlukile kwama-primers, ithempulethi DNA ne-MGCL2. Ukusabela ngakunye kwe-PCR kungadinga izimo ezihlukile zokusebenza kahle.

Sebenzisa ama-enzyme athembekile aphezulu: Uma ukunemba kubalulekile, cabanga ngokusebenzisa ukwethembeka okuphezulu kwe-DNA polymerase ukunciphisa amaphutha ngesikhathi sokukhuliswa.

Isixazululo: Yenza ukuhlolwa kokusebenzisa kahle ukuthola izimo ezinhle kakhulu zokusetha kwakho kwe-PCR ethile.

Ngokufigqiwe

Ukuxazulula inkinga aUkuhlaziywa kwe-PCRUngaba umsebenzi ovusana, kepha ukuqonda izinkinga ezijwayelekile nezixazululo zazo kungathuthukisa kakhulu isipiliyoni sakho se-PCR. Ngokuxazulula lezi zinkinga ezivamile, abacwaningi bangathuthukisa imiphumela ye-PCR futhi baqinisekise imiphumela ethembekile kuzinhlelo zokusebenza ze-molecular Biology. Ukulungiswa njalo, ukukhethwa ngokucophelela ama-reagents, nokwenza kahle kwezimo zokuphendula kuyizikhiye zokuhlaziywa kwe-PCR okuphumelelayo.