Ndikhulupirira kuti aliyense amakumana ndi zovuta zotere kapena zotere pochita machitidwe a PCR, koma ambiri aiwo amatha kugawidwa m'magulu awiri:

Kukulitsa pang'ono kwa template ya jini (kukulitsa);
Kuchulukitsitsa kwa jini kopanda cholinga.
Kugwiritsa ntchito zowonjezera ndi imodzi mwa njira zodziwika bwino zothetsera mavutowa.Nthawi zambiri ntchito ya zowonjezera imakhala ndi mbali ziwiri:
dongosolo lachiwiriza majini (kapangidwe kachiwiri);
Chepetsani kuyambika kosakhazikika.
Lero, mkonzi akufotokozerani mwachidule zowonjezera zomwe zimachitika mu PCR ndi ntchito zake.
Zowonjezera zomwe zimachepetsa dongosolo lachiwiri
sulfoxide(DMSO)
gene zitsanzoyokhala ndi GC yayikulu.Komabe, DMSO imachepetsanso kwambiri ntchito ya Taq polymerase.Chifukwa chake, aliyense ayenera kulinganiza kupezeka kwa template ndi ntchito za polymerase.Mkonzi akuwonetsa kuti mutha kuyesa mitundu yosiyanasiyana ya DSMO, monga kuchokera ku 2% mpaka 10%, kuti mupeze zomwe zikugwirizana ndi zomwe mumayesa.
Non-ionic zotsukira
Zotsukira zopanda ionic, monga 0.1-1% Triton X-100, Tween 20 kapena NP-40, nthawi zambiri zimachepetsa DNA yachiwiri.Ngakhale izi zitha kukulitsa kukulitsa kwa template ya jini, zingayambitsenso vuto la kukulitsa kosagwirizana kwenikweni.Chifukwa chake, zowonjezera izi zimagwira ntchito bwino pamachitidwe otsika a PCR opanda zinyalala, koma osati bwino pamachitidwe odetsedwa a PRC.Phindu lina la zotsukira zopanda ionic ndikuchepetsa kuipitsidwa kwa SDS.Nthawi zambiri pakupanga kwa DNA, SDS idzabweretsedwa ku sitepe ya PCR, yomwe imalepheretsa kwambiri ntchito ya polymerase.Chifukwa chake, kuwonjezera 0.5% Tween-20 kapena Tween-40 pakuchitako kumatha kusokoneza zotsatira zoyipa za SDS.
Betaine_
Betaine imatha kukulitsa kukulitsa kwa DNA pochepetsa mapangidwe achiwiri ndipo nthawi zambiri imakhala "chinsinsi" chowonjezera pa zida za PCR zamalonda.Ngati mukufuna kugwiritsa ntchito betaine, muyenera kuyika betaine kapena betaine mono-hydrate (Betaine kapena Betaine mono-hydrate), koma osati betaine hydrochloride (Betaine HCl), sinthani mpaka ndende yomaliza ya 1-1.7M.Betaine imatha kuthandizanso kuwongolera mwachindunji chifukwa imachotsa kudalira kwa DNA kusungunuka/kusintha kwa DNA.
Zowonjezera kuti muchepetse priming yosatchulika
Formamide
Formamide ndi chowonjezera cha PCR chomwe chimagwiritsidwa ntchito kwambiri.Ikhoza kuphatikiza ndi poyambira zazikulu ndi poyambira zazing'ono mu DNA, potero kuchepetsa kukhazikika kwa master DNA double helix ndikuchepetsa kutentha kwa DNA.Kuchuluka kwa formamide komwe kumagwiritsidwa ntchito poyeserera kwa PCR nthawi zambiri kumakhala 1% -5%.
Tetramethylammonium kloridi( TMAC)
Tetramethylammonium chloride imatha kukulitsa kuphatikizika kwa hybridization (kuphatikiza kusakanikirana) ndikuwonjezera kutentha kwa DNA.Chifukwa chake, TMAC imatha kuchotsa mapriming osakhazikika ndikuchepetsa kusokoneza kwa DNA ndi RNA.Ngati mugwiritsa ntchitozoyamba zowonongekamu zochita za PCR, kumbukirani kuwonjezera TMAC, yomwe imagwiritsidwa ntchito kwambiri pamagulu a 15-100mM.
Zina Zowonjezera Zowonjezera
Kuphatikiza pa mitundu iwiri ya zowonjezera zomwe tazitchula pamwambapa, pali zowonjezera zambiri zomwe zimachitika mu PCR, ngakhale zili ndi ntchito zosiyanasiyana, ndizofunikanso kwambiri.
Magnesium ion
Magnesium ion ndi cofactor (cofactor) yofunikira kwambiri ya polymerase, ndiye kuti, popanda magnesium ion, polymerase siyigwira ntchito.Komabe, ma magnesium ions ochulukirapo amathanso kukhudza mphamvu ya polymerase.Kuchuluka kwa ayoni a magnesium munjira iliyonse ya PCR kumasiyana.Chelating agents (monga EDTA kapena citrate), kuchuluka kwa dNTPs ndi mapuloteni onse zimakhudza kuchuluka kwa ayoni a magnesium.Chifukwa chake, ngati muli ndi zovuta pakuyesa kwanu kwa PCR, mutha kuyesa kusintha magawo osiyanasiyana a magnesium ion, mwachitsanzo, kuchokera ku 1.0 mpaka 4.0mM, ndi nthawi ya 0.5-1mM pakati.
Ndizofunikira kudziwa kuti kuzizira kowirikiza kangapo kungayambitse kukhazikika kwa magnesium chloride solution.Choncho, muyenera kusungunuka kwathunthu musanagwiritse ntchito, ndikusakaniza bwino musanagwiritse ntchito.
Bovine serum albumin(Bovine albumin, BSA)
Pakuyesa kwa chemistry ya molekyulu, seramu ya bovine albumin ndi chowonjezera chofala kwambiri, makamaka pakuletsa kugayidwa kwa enzyme ndi kuyesa kwa PCR.Mu machitidwe a PCR, BSA imathandiza kuchepetsa zonyansa monga phenolic compounds.Ndipo zimanenedwanso kuti zimatha kuchepetsa kumamatira kwa reactants ku khoma la test chubu.Mu machitidwe a PCR, nthawi zambiri kuchuluka kwa BSA kowonjezera kumatha kufika 0.8 mg/ml.
 
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PCR Hero