Ukubaluleka kokuLungiswa kokuSebenza kwe-Thermal Cycler

Iibhayisekile ezishushuzizixhobo eziyimfuneko kwicandelo lebhayoloji yemolekyuli kunye nophando lwemfuzo. Ezi zixhobo zidla ngokubizwa ngokuba yiPCR (ipolymerase chain reaction) ukuze kwandiswe ulandelelwano lweDNA, nto leyo evumela izazinzulu ukuba zenze imifuniselo eyahlukahlukeneyo ukusuka kwi-cloning ukuya kuhlalutyo lwembonakalo yemfuza. Nangona kunjalo, ukusebenza komjikelo we-thermal kuxhomekeke kakhulu kwi-calibration yayo, ngoko ke abaphandi kufuneka baqonde ukubaluleka kwale nkqubo.

Ulungelelwaniso yinkqubo yokulungelelanisa kunye nokuqinisekisa ukuchaneka kwemilinganiselo yesixhobo ngokuchasene nomgangatho owaziwayo. Kumjikelo we-thermal, oku kubandakanya ukuqinisekisa ukuba izicwangciso zobushushu zichanekile kwaye aziguquguquki kuyo yonke inkqubo yokukhwela ibhayisekile. Ukuchaneka kulawulo lobushushu kubalulekile, njengoko nokuphambuka okuncinci kunokukhokelela kumahluko omkhulu kwiziphumo zovavanyo lwe-PCR. Ngokomzekelo, ukuba iqondo lobushushu alifikelelwanga, imisonto ye-DNA isenokungohlukani kakuhle, nto leyo ebangela ukuba inganyuswa kakuhle. Ngokufanayo, ukuba iqondo lobushushu le-annealing lisezantsi kakhulu okanye liphezulu kakhulu, kunokukhokelela ekubopheni okungangxengwanga okanye ukunqongophala ngokupheleleyo kokuzibophelela, ekugqibeleni kubeke esichengeni isidima sovavanyo.

Esinye sezizathu eziphambili zokuba kutheni ulungelelwaniso lubaluleke kakhulu kwiibhayisekile ezishushu yimpembelelo enayo ekwenziweni kwakhona. Kuphando lwezenzululwazi, ukuveliswa kwakhona lilitye lembombo lokuthembeka. Ukuba i-thermal cycler ayilinganisi ngokuchanekileyo, iziphumo ezifunyenwe kwiimvavanyo ezahlukeneyo zinokwahluka, okwenza kube nzima ukuphinda iziphumo zophando. Oku kungahambelani kunokukhokelela kwizigqibo eziphosakeleyo kunye nemithombo echithwayo, ebeka esichengeni ukuqinisekiswa okupheleleyo kophando. Ukulinganisa rhoqo kuqinisekisa ukuba i-thermal cycler isebenza ngaphakathi kweeparamitha ezichaziweyo, ngaloo ndlela ukwandisa ukuthembeka kweziphumo zakho.

Ngaphezu koko, ukubaluleka kokulinganisa akukona nje ukuchaneka kwemimiselo yeqondo lokushisa, kodwa nakwindlela efanayo yokusabalalisa ukushisa ngaphakathi kwe-thermal cycler. Isixhobo esilungelelaniswe kakuhle kufuneka sikwazi ukubonelela ngamaqondo obushushu ahambelanayo kuwo onke amaqula kwipleyiti ye-multiwell. Ukuhluka kobushushu kunokukhokelela ekuhlukeni kwimilinganiselo yokukhulisa, enokuchaphazela iziphumo kwaye ekugqibeleni isiphumo esipheleleyo sovavanyo. Ngokulinganisa umjikelo we-thermal, abaphandi banokuqinisekisa ukuba zonke iisampuli ziphantsi kweemeko ezifanayo zokushisa, ngaloo ndlela ziphucula umgangatho wedatha.

Ukongeza ekuphuculeni ukuchaneka kunye nokuphindaphinda, ukulinganisa rhoqo umjikelo wakho we-thermal kunokwandisa ubomi besixhobo. Ngokuhamba kwexesha, izinto ezingaphakathi kwi-thermal cycler zinokuphelelwa okanye zingasebenzi kakuhle, nto leyo ekhokelela kwiimpazamo ezinokwenzeka. Ngokulinganisa izixhobo rhoqo, abaphandi banokuchonga kwaye basombulule imiba ngaphambi kokuba ibe yingxaki enkulu, baqinisekise ukuba i-thermal cycler ihlala ikwindlela yokusebenza efanelekileyo. Le ndlela isebenzayo yokulinganisa ayigcini nje kuphela iindleko ezinxulumene nokulungiswa okanye ukutshintshwa, kodwa inciphisa ixesha lokunciphisa elebhu.

Ngamafutshane, ulungelelwaniso lweiibhayisekile ezishushungumba osisiseko wokuqinisekisa ukusebenza kwawo kunye nokuthembeka kuphando lwezenzululwazi. Ukulawula ubushushu obuchanekileyo kunye nokufana kubalulekile kwimpumelelo ye-PCR kunye nezinye iimvavanyo ezixhomekeke kubushushu. Ngokwenza ukulinganisa rhoqo kube yinto ephambili, abaphandi banokuphucula ukuphindaphinda kweziphumo, ukugcina imfezeko yeziphumo zabo, kunye nokwandisa ubomi bezixhobo zabo. Njengoko icandelo lebhayoloji yemolekyuli liqhubeka nokuhambela phambili, ukubaluleka kokulinganisa i-thermal cycler calibration kuya kuqhubeka nokuba yinto ephambili ekuqhubeni inkqubela phambili yesayensi kunye nokutsha.