Kwimveliso yeeflange ezinkulu, kukho izinto ezininzi ezichaphazela ukusebenza kweeflange ezinkulu. Apha ngezantsi sithetha izinto ezininzi eziqhelekileyo, eyokuqala liqondo lobushushu lobushushu (iqondo lobushushu le-annealing yiparameter yobushushu xa i-primer kunye netemplate zidityanisiwe, xa i-50% ye-primers i-Iqondo lobushushu apho ulandelelwano oluhambelanayo lubonakaliswa njenge-double-- I-annealing DNA molekyuli yinto ebalulekileyo echaphazela ubunjani be-PCR Phantsi kweemeko ezifanelekileyo, iqondo lobushushu le-annealing lisezantsi ngokwaneleyo ukuze kuqinisekiswe ukuba ii-primers zifakwe ngokufanelekileyo kulandelelwano olufunwayo, ngelixa liphezulu ngokwaneleyo ukunciphisa ukubophelela okungangqalanga iqondo lobushushu ukusuka 55 ° C ukuya 70 ° C. Iqondo lobushushu annealing ngokubanzi iseti 5 ° C ngaphantsi kwe Tm primer.

Ukuba ubushushu obuchaziweyo bufikelelwe. Ubushushu be-anealing bufikelela kubushushu obufunekayo. Unyango olukhulu lweflange luqhutywa ngokubanzi ngonyango oluluqilima lobushushu, olubizwa ngokuqhelekileyo ngokuthi "i-annealing", kunye noluhlu lobushushu yi-1040 ~ 1120 °C (umgangatho waseJapan). Unokujonga kwakhona ngomngxuma wokujonga kwiziko lomlilo. Ukufakelwa kwe-flange enkulu kwindawo yokuthungatha kufuneka ibe kwindawo ye-incandescent, kodwa akukho nto ithambileyo kunye ne-sagging.

Ilandelwa kukutywinwa komzimba wesithando somlilo (iqokobhe langaphandle lidityaniswe ngepleyiti yentsimbi kunye nentsimbi yecandelo, itroli idityaniswe ngentsimbi kunye nepleyiti yentsimbi, itroli idlulisa unxibelelwano oluthambileyo kunye nelinen yesithando somlilo kunye nendlela yokutywinwa kwesanti ukunciphisa ubushushu bemitha. kunye nokulahlekelwa kwe-convection, kunye nokuqinisekisa ngokufanelekileyo ukutywinwa komzimba wesithando somlilo ), isithando somlilo esikhanyayo kufuneka sivalwe kwaye sihlukaniswe ngaphandle emoyeni; usebenzisa i-hydrogen njengegesi ekhuselayo, i-port enye yokukhupha ivuliwe (isetyenziselwa ukutshisa i-hydrogen ekhutshiweyo). Indlela yokuhlola ingasetyenziswa kwizithuba zamalungu esithando somlilo ngamanzi anesepha ukubona ukuba irhasi iyasebenza. Indawo elula kakhulu yokuqhuba igesi yindawo apho isithando somlilo singena kumbhobho kunye nendawo apho umbhobho ukhutshwa khona. Umsesane wokutywina kule ndawo ulungele ukunxiba. Soloko ujonga rhoqo ukuze utshintshe.

Okwesibini, uxinzelelo lwegesi olukhuselayo, ukwenzela ukuthintela ukuvela kwe-micro-leakage, igesi yokukhusela iziko kufuneka igcine uxinzelelo oluthile oluhle, ukuba igesi yokukhusela i-hydrogen, ifuna ngokubanzi ngaphezu kwe-20kBar. I-Annealing atmosphere: Ngokubanzi, i-hydrogen esulungekileyo isetyenziswa njenge-atmosfera yomoya, kwaye ukucoceka kwe-atmosfera kukhethwa ngama-99.99% nangaphezulu. Ukuba enye indawo ye-atmosfera yigesi engasebenziyo, ukucoceka kunokuba ngaphantsi, kodwa akufanele kuqulethe i-oxygen eninzi okanye umphunga wamanzi.