Ini ndinotenda kuti munhu wese anogara achisangana nematambudziko akadai kana akadaro pakuita PCR maitiro, asi mazhinji acho anogona kuiswa mumatambudziko makuru maviri:

Kuwedzera kudiki kweiyo gene template (kukudza);
Yakawandisa isiri-chinangwa gene amplification.
Kushandisa additives ndiyo imwe yenzira dzakajairika kugadzirisa matambudziko aya.Kazhinji basa rekuwedzera rine zvinhu zviviri:
chimiro chechipiriyemajini (secondary structure);
Deredza priming isiri-chaiyo.
Nhasi, mupepeti achasuma kwauri muchidimbu izvo zvakajairika zvinowedzera muPCR maitiro uye mabasa avo.
Zviwedzerwa zvinoderedza chimiro chechipiri
sulfoxide(DMSO)
gene samplesine yakakwira GC yemukati.Nekudaro, DMSO zvakare inoderedza zvakanyanya Taq polymerase chiitiko.Naizvozvo, munhu wese anofanirwa kuenzanisa kuwanikwa kwetemplate uye chiitiko chepolymerase.Mupepeti anoratidza kuti unogona kuedza zvakasiyana-siyana zveDSMO, zvakadai kubva pa2% kusvika ku10%, kuti uwane kusungirirwa kunoenderana nekuedza kwako.
Non-ionic detergents
Non-ionic detergents, zvakadai se0.1-1% Triton X-100, Tween 20 kana NP-40, kazhinji inoderedza DNA yechipiri chimiro.Kunyangwe izvi zvichigona kuwedzera kukwidziridzwa kweiyo template gene, zvinozokonzeresa dambudziko rekusina-chaiyo amplification.Saka, izvi zvinowedzera zvinoshanda zvakanaka kune yakaderera-goho PCR maitiro asina marara, asi kwete zvakanyanya kune isina kuchena PRC maitiro.Imwe bhenefiti yeasiri-ionic detergents ndiko kudzikiswa kweSDS kusvibiswa.Kazhinji panguva yekutorwa kweDNA, SDS ichaunzwa kuPCR nhanho, iyo inodzivirira zvakanyanya basa repolymerase.Naizvozvo, kuwedzera 0.5% Tween-20 kana Tween-40 kune maitiro anogona kuderedza mhedzisiro yakaipa yeSDS.
Betaine_
Betaine inogona kuvandudza DNA nekudzikisa kuumbwa kwechipiri chimiro uye kazhinji "chakavanzika" chekuwedzera kune ekutengesa PCR kits.Kana iwe uchida kushandisa betaine, unofanira kuisa betaine kana betaine mono-hydrate (Betaine kana Betaine mono-hydrate), asi kwete betaine hydrochloride (Betaine HCl), gadzirisa kusvika kuhuwandu hwekupedzisira hwe1-1.7M.Betaine inogonawo kubatsira kuvandudza hunhu nekuti inobvisa base base pair kutsamira kweDNA kunyunguduka / DNA denaturation.
Additives kuderedza zvisina-chaiyo priming
Formamide
Formamide inowanzoshandiswa organic PCR yekuwedzera.Inogona kusanganisa negororo hombe uye diki groove muDNA, nekudaro ichidzikisa kugadzikana kwatenzi DNA kaviri helix uye kudzikisa kupisa kwekunyungudika kweDNA.Iko kusanganiswa kweformamide inoshandiswa mukuedzwa kwePCR kunowanzo 1% -5%.
Tetramethylammonium chloride( TMAC)
Tetramethylammonium chloride inogona kuwedzera iyo chaiyo yekusanganiswa (hybridization chaiyo) uye kuwedzera tembiricha yekunyungudika yeDNA.Nekudaro, TMAC inogona kubvisa zvisiri-chaiyo priming uye kuderedza kusasungirirwa kweDNA neRNA.Kana ukashandisadegenerate primersmuPCR reaction, yeuka kuwedzera TMAC, iyo inowanzoshandiswa pamusangano we15-100mM.
Zvimwe Zvimwe Zvinowedzerwa
Pamusoro pezvikamu zviviri zvezvinowedzera zvataurwa pamusoro apa, kune zvakawanda zvinowanzoitika muPCR maitiro, kunyange zvazvo ane mabasa akasiyana, akakoshawo.
Magnesium ion
Magnesium ion chinhu chakakosha cofactor (cofactor) yepolymerase, kureva kuti, pasina magnesium ion, polymerase haishande.Nekudaro, yakawandisa magnesium ion inogona kukanganisa kushanda kwepolymerase.Iko kusanganiswa kwe magnesium ions mune yega PCR maitiro ichasiyana.Chelating agents (zvakadai seEDTA kana citrate), kuwanda kwe dNTPs uye mapuroteni zvose zvinokanganisa kusanganiswa kwe magnesium ions.Saka, kana uine matambudziko nekuedza kwako kwePCR, unogona kuedza kushandura magnesium ion concentrations dzakasiyana, semuenzaniso, kubva pa1.0 kusvika 4.0mM, paine nguva ye0.5–1mM pakati.
Izvo zvakakosha kuti ticherechedze kuti kuwanda kwechando-kunyungudika kutenderera kunogona kutungamira mukusungwa stratification ye magnesium chloride mhinduro.Naizvozvo, unofanira kuinyungudutsa zvachose usati washandisa, woisanganisa zvakanaka usati waishandisa.
Bovine serum albumin(Bovine albumin, BSA)
Muchiyedzo chemakemikari makemikari, bovine serum albumin chinhu chakajairika chekuwedzera, kunyanya mukudzora enzyme digestion uye PCR kuyedza.Mukuita kwePCR, BSA inobatsira mukudzikisa zvinosvibisa senge phenolic compounds.Uye zvakare inonzi inogona kuderedza kusungirirwa kwemareactants kumadziro e test tube.Muchiitiko chePCR, kazhinji huwandu hweBSA hwakawedzerwa hunogona kusvika 0.8 mg / ml.
 
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