Ekukhiqizeni ama-flanges amakhulu, kunezici eziningi ezithinta ukusebenza kwama-flange amakhulu. Ngezansi sisho izici ezimbalwa ezivamile, eyokuqala yizinga lokushisa lokukhipha isisu (izinga lokushisa le-annealing ipharamitha yezinga lokushisa lapho i-primer kanye nesifanekiso kuhlanganiswa, lapho ama-50% weziqalisi kukhona Izinga lokushisa lapho ukulandelana okuhambisanayo kuvezwa khona njengokuphindwe kabili- I-molecule ye-DNA exhumekile iyisici esibalulekile esithinta ukucaciswa kwe-PCR Ngaphansi kwezimo ezikahle, izinga lokushisa le-anneal liphansi ngokwanele ukuze kuqinisekiswe ukuthi ama-primer axhumene ngempumelelo nokulandelana okufunayo, kuyilapho ephakeme ngokwanele ukunciphisa ukubopha okungaqondile Izinga lokushisa lisuka ku-55 ° C kuya ku-70 ° C. Izinga lokushisa le-anneal ngokuvamile libekwe ngo-5 ° C ngaphansi kune-Tm ye-primer.

Ukuthi izinga lokushisa elishiwo lifinyelelwe yini. Izinga lokushisa le-annealing lifinyelela izinga lokushisa elidingekayo. Ukwelashwa kwe-flange ngezinga elikhulu ngokuvamile kwenziwa ukwelashwa kokushisa kwesisombululo esiqinile, okuvame ukubizwa ngokuthi "i-annealing", futhi izinga lokushisa lingu-1040 ~ 1120 °C (okujwayelekile kwaseJapane). Ungabuka futhi ngembobo yokubuka yesithando somlilo. Ukufakwa kwe-flange enkulu endaweni ye-anneal kufanele kube kusimo se-incandescent, kodwa akukho ukuthambisa nokuhlehla.

Kulandelwa ukubekwa uphawu kwesithando somlilo (igobolondo langaphandle lishiselwe ngensimbi yensimbi kanye nensimbi yesigaba, i-trolley ishiselwe ngensimbi ne-steel plate, i-trolley idlulisa ukuthintana okuthambile nolayini lwesithando somlilo kanye nendlela yokuvala isihlabathi ukuze kuncishiswe imisebe yokushisa. kanye nokulahlekelwa kwe-convection, futhi uqinisekise ngokuphumelelayo ukubekwa uphawu kwesithando somlilo ), isithando somlilo esikhanyayo kufanele sivalwe futhi sihlukaniswe nomoya wangaphandle; usebenzisa i-hydrogen njengegesi evikelayo, imbobo yokukhipha umoya eyodwa kuphela evuliwe (esetshenziselwa ukuthungela i-hydrogen ekhishiwe). Indlela yokuhlola ingasetshenziswa ezikhaleni zamalunga esithando somlilo esinamanzi anensipho ukuze kubonakale ukuthi igesi iyasebenza yini. Indawo elula kakhulu yokuqhuba igesi yindawo lapho isithando somlilo singena khona epayipini nendawo lapho ipayipi ikhishwa khona. Indandatho yokuvala kule ndawo ijwayele ukugqoka. Njalo hlola njalo ukuze ushintshe.

Okwesibili, ingcindezi yegesi evikelayo, ukuze kuvinjwe ukuvela kokuvuza okuncane, igesi yokuvikela isithando somlilo kufanele igcine ukucindezela okuthile okuhle, uma kuyigesi yokuvikela i-hydrogen, ngokuvamile idinga okungaphezu kuka-20kBar. I-Annealing atmosphere: Ngokuvamile, i-hydrogen ehlanzekile isetshenziswa njengomkhathi oshisayo, futhi ukuhlanzeka komkhathi kungcono kube ngu-99.99% noma ngaphezulu. Uma enye ingxenye yomkhathi kuyigesi engasebenzi, ukuhlanzeka kungaba ngaphansi, kodwa akumele kuqukathe umoya-mpilo omningi noma umhwamuko wamanzi.