Ungawukhetha kanjani umjikelezo oshisayo olungile ngezidingo zakho zocwaningo

Amabhayisikili ashisayozingamathuluzi abalulekile uma kukhulunywa ngebhayoloji yamangqamuzana nocwaningo lwezakhi zofuzo. Eyaziwa nangokuthi umshini we-PCR (polymerase chain reaction), le divayisi ibalulekile ekukhuliseni i-DNA, iyenze ibe yitshe legumbi lezinhlelo zokusebenza ezihlukahlukene ezihlanganisa ukuhlanganisa, ukulandelana nokuhlaziya isisho sezakhi zofuzo. Kodwa-ke, kunezinketho eziningi emakethe kangangokuthi ukukhetha umjikelezo oshisayo olungile wezidingo zakho zocwaningo kungaba umsebenzi onzima. Nazi ezinye izici ezibalulekile okufanele uzicabangele lapho wenza ukukhetha kwakho.

1. Qonda izidingo zakho zocwaningo

Ngaphambi kokungena ekucacisweni kwamabhayisikili ashisayo ahlukene, kubalulekile ukuhlola izidingo zakho zocwaningo. Cabangela uhlobo lokuhlola ozolwenza. Ingabe usebenzisa i-PCR ejwayelekile, i-quantitative PCR (qPCR), noma uhlelo lokusebenza olusebenzisa kakhulu? Ngalunye lwalezi zinhlelo zokusebenza lungase ludinge izici ezihlukile namandla we-thermal cycler.

2. Izinga Lokushisa kanye Nokufanayo

Izinga lokushisa lomjikelezo oshisayo liyisici esibalulekile. Amaphrothokholi amaningi e-PCR adinga isinyathelo sokuguqula isimo cishe sibe ngu-94-98°C, isinyathelo sokukhipha isisu ku-50-65°C, kanye nesinyathelo sokunwetshwa kokuthi 72°C. Qiniseka ukuthi umjikelezo oshisayo owukhethayo uyakwazi ukuphatha lawa mazinga okushisa nokuthi izinga lokushisa lisatshalaliswa ngokulinganayo kuyo yonke imojuli. Ukungafani kahle kwezinga lokushisa kungathinta ucwaningo lwakho ngokubangela imiphumela engahambisani.

3. Ifomethi yokuvimba nomthamo

Amabhayisekili ashisayo afika ngamafomethi ahlukahlukene, kufaka phakathi amapuleti anemithombo engama-96, amapuleti emithombo engama-384, kanye namapuleti emithombo engu-1536. Ukukhethwa kwefomethi yebhulokhi kufanele kufane nezidingo zakho zokuphuma. Uma wenza izivivinyo ze-high-throughput, ungase udinge ifomethi yebhulokhi enkulu. Ngokuphambene, ekuhlolweni kwezikali ezincane, ipuleti lemithombo engu-96 lingase lanele. Ukwengeza, cabanga ukuthi uyawadinga yini amamojula ashintshekayo kumafomethi ahlukene, njengoba lokhu kungakhuphula ukuguquguquka kocwaningo lwakho.

4. Isivinini nokusebenza kahle

Esimweni sanamuhla socwaningo olusheshayo, isikhathi siwumongo. Bheka ibhayisikili elishisayo elinamandla okufudumeza okusheshayo namandla okupholisa. Amanye amamodeli athuthukile angaqeda umjikelezo we-PCR ngaphansi kwemizuzu engama-30, asheshise kakhulu ukuhamba kwakho komsebenzi. Ngaphezu kwalokho, izici ezifana nemodi esheshayo noma amazinga okushisa asheshayo akhuphula ukusebenza kahle, okukuvumela ukuthi ucubungule amasampuli amaningi ngesikhathi esincane.

5. Isixhumi esibonakalayo somsebenzisi neSoftware

Isixhumi esibonakalayo esisebenziseka kalula sibalulekile ekusebenzeni kahle. Bheka ibhayisikili elishisayo elinesikrini sokuthinta esinembile, izinketho ezilula zokuhlela, kanye nemithetho elandelwayo esethwe ngaphambilini. Amamodeli athuthukile angase futhi eze nesofthiwe evumela ukuqapha kwesikhathi sangempela nokuhlaziywa kwedatha, okuzuzisa kakhulu izinhlelo zokusebenza ze-qPCR. Qiniseka ukuthi isofthiwe iyahambisana namasistimu akho akhona futhi ingaphatha okuphumayo kwedatha okudingayo.

6. Ukucatshangelwa kwesabelomali

Amabhayisikili ashisayo ahluka kakhulu ngentengo, ngakho-ke kubalulekile ukuthi ube nesabelomali ngaphambi kokuthi uqale ukuyithenga. Nakuba kungase kulinge ukuhamba nenketho eshibhe kakhulu, cabanga ngenani lesikhathi eside lokutshala imali emshinini wekhwalithi ephezulu ohlangabezana nezidingo zakho zocwaningo. Ungacabangi nje kuphela inani lokuqala lokuthenga, kodwa futhi nezindleko zokusebenziseka, ukugcinwa, kanye nokuthuthukiswa okungenzeka.

7. Ukusekela Umkhiqizi kanye Newaranti

Okokugcina, cabangela izinga lokusekela kanye newaranti enikezwe umenzi. Ibhayisikili elishisayo elithembekile kufanele linikeze iwaranti ebanzi futhi libe nokusekelwa kwamakhasimende ekuxazululeni izinkinga nokunakekela. Lokhu kongela isikhathi nezinsiza ngokuhamba kwesikhathi.

Ekuphetheni

Ukukhetha okulungileumjikelezo oshisayongezidingo zakho zocwaningo yisinqumo esibucayi esingaba nomthelela empumelelweni yocwaningo lwakho. Ngokucubungula ngokucophelela izidingo zakho ezithile, izinga lokushisa, ifomethi yemojuli, isivinini, ukusebenzelana komsebenzisi, isabelomali, nokusekelwa komkhiqizi, ungenza ukukhetha unolwazi oluzothuthukisa amakhono akho ocwaningo futhi uthole imiphumela enokwethenjelwa. Ukutshala isikhathi kule nqubo yokukhetha ekugcineni kuzokhokha ngekhwalithi nokusebenza kahle komsebenzi wakho wesayensi.