Ke lumela hore motho e mong le e mong o tla lula a kopana le mathata a joalo kapa a joalo ha a etsa karabelo ea PCR, empa boholo ba bona bo ka aroloa ka mathata a mabeli a mantlha:

Kholiso e nyane haholo ea template ea liphatsa tsa lefutso (amplification);
Keketseho e ngata haholo ea mofuta oo e seng oa sepheo.
Ho sebelisa li-additives ke e 'ngoe ea mekhoa e tloaelehileng ea ho rarolla mathata ana.Hangata karolo ea li-additives e na le likarolo tse peli:
sebopeho sa bobediea liphatsa tsa lefutso (sebopeho sa bobeli);
Fokotsa lihlahisoa tse sa tloaelehang.
Kajeno, mohlophisi o tla u tsebisa ka bokhutšoanyane li-additives tse tloaelehileng liketsong tsa PCR le mesebetsi ea tsona.
Li-additives tse fokotsang sebopeho sa bobeli
sulfoxide(DMSO)
mehlala ea liphatsa tsa lefutsoe nang le GC e phahameng.Leha ho le joalo, DMSO e boetse e fokotsa haholo ts'ebetso ea Taq polymerase.Ka hona, motho e mong le e mong o tlameha ho leka-lekanya phihlello ea template le ts'ebetso ea polymerase.Mohlophisi o fana ka maikutlo a hore u ka leka likhakanyo tse fapaneng tsa DSMO, joalo ka ho tloha ho 2% ho isa ho 10%, ho fumana mohopolo o lumellanang le liteko tsa hau.
Lisebelisoa tse se nang ionic
Li-detergent tse se nang ionic, tse kang 0.1-1% Triton X-100, Tween 20 kapa NP-40, hangata li fokotsa sebopeho sa bobeli sa DNA.Leha sena se ka eketsa ho holisoa ha mofuta oa template, se tla baka le bothata ba ho holisa ka mokhoa o sa tobang.Ka hona, li-additives tsena li sebetsa hantle bakeng sa karabelo ea PCR ea kotulo e tlase ntle le maloanlahla, empa e seng hantle bakeng sa karabelo e batlang e sa hloeka ea PRC.Molemo o mong oa li-detergent tse seng tsa ionic ke phokotso ea tšilafalo ea SDS.Hangata nakong ea ts'ebetso ea ho ntša DNA, SDS e tla tlisoa mohato oa PCR, o thibelang haholo mosebetsi oa polymerase.Ka hona, ho eketsa 0.5% Tween-20 kapa Tween-40 karabelong ho ka fokotsa litlamorao tse mpe tsa SDS.
Betaine_
Betaine e ka ntlafatsa matlafatso ea DNA ka ho fokotsa sebopeho sa sebopeho sa bobeli mme ka kakaretso ke tlatsetso ea "sephiri" ho lisebelisoa tsa PCR tsa khoebo.Haeba u batla ho sebelisa betaine, u lokela ho beha betaine kapa betaine mono-hydrate (Betaine kapa Betaine mono-hydrate), empa eseng betaine hydrochloride (Betaine HCl), ikamahanya le maemo a ho qetela a 1-1.7M.Betaine e ka boela ea thusa ho ntlafatsa ho khetheha hobane e felisa ho itšetleha ka sebopeho sa motheo sa DNA melting/DNA denaturation.
Li-additives tsa ho fokotsa li-priming tse sa tobang
Formamide
Formamide ke motsoako o sebelisoang hangata oa PCR.E ka kopana le groove e kholo le groove e nyane ho DNA, ka hona ho fokotsa botsitso ba master DNA double helix le ho theola mocheso o qhibilihang oa DNA.Motsoako oa formamide o sebelisitsoeng litekong tsa PCR hangata ke 1% -5%.
Tetramethylammonium chloride( TMAC)
Tetramethylammonium chloride e ka eketsa ho hlaka ha hybridization (ka ho khetheha ho nyalisitsoeng) le ho eketsa mocheso o qhibilihang oa DNA.Kahoo, TMAC e ka tlosa priming e sa tobang le ho fokotsa ho kopanya hampe ha DNA le RNA.Haeba u sebelisali-primers tse senyehilengka karabelo ea PCR, hopola ho eketsa TMAC, e sebelisoang hangata ka bongata ba 15-100mM.
Li-additives tse ling tse tloaelehileng
Ho phaella ho lihlopha tse peli tsa li-additives tse boletsoeng ka holimo, ho na le li-additives tse ngata tse tloaelehileng liketsong tsa PCR, le hoja li na le mesebetsi e sa tšoaneng, hape li bohlokoa haholo.
Magnesium ion
Magnesium ion ke cofactor ea bohlokoa (cofactor) ea polymerase, ke hore, ntle le ion magnesium, polymerase ha e sebetse.Leha ho le joalo, li-ion tse ngata tsa magnesium le tsona li ka ama katleho ea polymerase.Khatello ea li-ion tsa magnesium karabelong e 'ngoe le e' ngoe ea PCR e tla fapana.Chelating agents (tse kang EDTA kapa citrate), mahloriso a dNTP le liprotheine kaofela li ama bongata ba li-ion tsa magnesium.Kahoo, haeba u na le mathata ka teko ea hau ea PCR, u ka leka ho fetola metsoako e fapaneng ea magnesium ion, mohlala, ho tloha 1.0 ho ea ho 4.0mM, ka nako ea 0.5-1mM pakeng tsa.
Ho bohlokoa ho ela hloko hore lipotoloho tse ngata tsa ho qhibiliha ha leqhoa li ka lebisa tlhokomelong ea stratification ea tharollo ea magnesium chloride.Ka hona, o tlameha ho e qhaqha ka ho feletseng pele o sebelisoa, 'me u e kopanye hantle pele u e sebelisa.
Bovine serum albumin(Bovine albumin, BSA)
Litekong tsa k'hemistri ea limolek'hule, albumin ea serum ea bovine ke tlatsetso e tloaelehileng haholo, haholo-holo litekong tse thibelang tšilo ea enzyme le liteko tsa PCR.Lits'ebetsong tsa PCR, BSA e thusa ho fokotsa litšila tse kang metsoako ea phenolic.Hape ho boleloa hore e ka fokotsa ho khomarela ha li-reactants leboteng la tube ea teko.Karabelo ea PCR, hangata khatello ea BSA e kentsoeng e ka fihla ho 0.8 mg/ml.
 
Lihlahisoa tse Amanang:
Mohale oa PCR(ka dae)
Mohale oa PCR