Ho Ntlafatsa Bokhoni ba PCR ka ho Sebelisa Li-cyclers tsa Thermal tse tsoetseng pele

Polymerase chain reaction (PCR) ke mokhoa oa mantlha oa baeloji ea limolek'hule 'me o sebelisoa haholo ho holisa tatelano ea DNA. Ho sebetsa hantle le ho nepahala ha PCR ho angoa haholo ke sebaesekele sa mocheso se sebelisoang ts'ebetsong. Libaesekele tse tsoetseng pele tsa mocheso li bapala karolo ea bohlokoa ho ntlafatseng ts'ebetso ea PCR, ho fana ka taolo e nepahetseng ea mocheso, litefiso tse potlakileng tsa ho futhumatsa le ho pholisa, le bokhoni bo tsoetseng pele ba mananeo.

E 'ngoe ea likarolo tsa bohlokoa tsa e tsoetseng pelelibaesekele tse chesangke taolo e nepahetseng ea mocheso. Ho boloka lithempereichara tse khethehileng bakeng sa mehato ea denaturation, annealing, le katoloso ho bohlokoa bakeng sa ho holisa PCR ka katleho. Sebaesekele se tsoetseng pele sa mocheso se tiisa kabo e ts'oanang le e nepahetseng ea mocheso ka har'a liliba tsohle tsa lisampole, se fokotsa ho fapana ha matla a ho holisa le ho fokotsa monyetla oa ho holisa ntle le ho khetheha.

Likhahla tse potlakileng tsa ho futhumatsa le ho pholisa ke karolo e 'ngoe ea bohlokoa ea libaesekele tse tsoetseng pele tsa mocheso. Lisebelisoa tsena li na le theknoloji e thehiloeng ho Peltier e ka fetohang ka potlako pakeng tsa mehato e fapaneng ea mocheso. Paesekele ena e potlakileng ea mocheso ha e boloke nako feela empa e boetse e fokotsa kotsi ea sebopeho sa "primer-dimer" le ho holisa ka mokhoa o sa tobang, ka tsela eo e eketsa boits'oaro le bokhoni ba PCR.

Ntle le moo, libaesekele tse tsoetseng pele tsa mocheso li fana ka bokhoni ba mananeo a tsoetseng pele, a lumellang basebelisi ho etsa liprothokholo tsa PCR ho latela litlhoko tsa bona tsa liteko. Lisebelisoa tsena li fana ka monyetla oa ho theha gradient PCR, ho landing PCR, le liprothokholo tse ling tse ikhethang, tse nolofalletsang ho ntlafatsa maemo a PCR bakeng sa li-primer sets le litempele tse fapaneng. Ntle le moo, libaesekele tse ling tse tsoetseng pele tsa mocheso li na le likhokahano tse hlakileng tsa software tse nolofatsang moralo oa protocol le tlhahlobo ea data, ka hona e ntlafatsa katleho ea liteko.

Ntle le likarolo tsena, libaesekele tse ling tse tsoetseng pele tsa mocheso li fana ka mahlale a morao-rao joalo ka likoahelo tse futhumetseng tse thibelang ho koaleha le ho tsoa mouoane nakong ea libaesekele tsa PCR, ho netefatsa maemo a ts'ebetso a tsitsitseng le ho fokotsa tahlehelo ea sampole. Tse ling li ka kenyelletsa ts'ebetso ea gradient e ka ntlafatsang lithemparetjha tsa annealing bakeng sa disampole tse ngata ka nako e le 'ngoe, ho ntlafatsa ts'ebetso ea PCR le ho ts'epahala.

Bohlokoa ba ho sebelisa sebaesekele se tsoetseng pele sa mocheso ho ntlafatsa ts'ebetso ea PCR bo ke ke ba feteletsoa. Lisebelisoa tsena ha li nolofatse ts'ebetso ea PCR feela empa hape li thusa ho ntlafatsa ho ikatisa le ho nepahala ha liphetho tsa liteko. Ka ho fana ka taolo e nepahetseng ea mocheso, ho palama libaesekele tse potlakileng, le bokhoni ba mananeo a tsoetseng pele, libaesekele tse tsoetseng pele tsa mocheso li thusa bafuputsi ho fihlela ntlafatso e matla, e sebetsang ea PCR bakeng sa lits'ebetso tse fapaneng, ho kenyelletsa tlhahlobo ea polelo ea liphatsa tsa lefutso, genotyping le cloning.

Qetellong, e tsoetseng pelelibaesekele tse chesangbapala karolo ea bohlokoa ho ntlafatseng ts'ebetso ea PCR. Taolo ea eona e nepahetseng ea mocheso, likhahla tse potlakileng tsa ho futhumatsa le ho pholisa, le bokhoni ba mananeo a tsoetseng pele li thusa ho ntlafatsa ho nepahala, ho hlaka, le ho ikatisa ha PCR amplification. Bafuputsi ba ka rua molemo o moholo ka ho sebelisa libaesekele tse tsoetseng pele tsa mocheso litekong tsa baeloji ea limolek'hule, e leng se qetellang se lebisa ho litšibollo tse tšepahalang le tse nang le temohisiso tsa mahlale.