Zomwe zimachitika mu PCR zimachitika

Pa nthawi ya PCR, zina zomwe zikuchitika zimakumana nthawi zambiri.
Chifukwa cha chidwi chachikulu cha PCR, kuipitsidwa kumawerengedwa kuti ndi chimodzi mwazinthu zofunikira kwambiri zomwe zimakhudza zotsatira za PCR ndipo zimatha kupanga zotsatira zabwino.
Cholinga chofananira ndi magawo osiyanasiyana omwe amatsogolera ku zotsatira zoyipa. Ngati gawo limodzi kapena zingapo zofunikira za PCR yosakaniza kapena njira zowonjezera zomwe zimalephereka kapena kusokonezedwa ndi vuto la diagnastic ikhoza kulephereka. Izi zimatha kubweretsa kuchepa kwamphamvu komanso zotsatira zoyipa zabodza.
Kuphatikiza pa zoletsa, kutayika kwa chandamale cha nuclec acid kuchitika chifukwa chotumiza ndi / kapena malo osungira asanakonzekere. Makamaka, kutentha kwambiri kapena kusungirako kokwanira kumatha kubweretsa kuwonongeka kwa maselo ndi ma acid acid. Selo ndi minofu ya minofu ndi paraffin yophatikizidwa ndi zifukwa zodziwika bwino za DNA komanso vuto lopitilira (onani 2 ndi 2). Muzochitika izi, ngakhale kudzipatula koyenera komanso kuyeretsa sikungathandize.

Chithunzi 1 | Zotsatira za Undebolization pa DNA Kukhulupirika
Agarose gel electrophoresis adawonetsa kuti mtundu wa DNA udatalikirana ndi zigawo za parafini za ma autopries osiyanasiyana. DNA ya kutalika kosiyanasiyana kudalipo m'manda otengera njira yosinthira. DNA idasungidwa pokhapokha ngati ziwonetsero za anthu achisanu komanso mu osalowerera ndale. Kugwiritsa ntchito bouin mwamphamvu kapena osafunikira, mawonekedwe a acid okhala ndi acid omwe ali ndi mwayi wotayika kwambiri kwa DNA. Gawo lotsala limagawidwa kwambiri.
Kumanzere, kutalika kwa zidutswa kumafotokozedwa mu kilobase awiriawiri (KBP)

Chithunzi 2 | Kutayika kwa Umphumphu wa Nucleic acid
. Kaphatikizidwe wa DNA idzachitikabe kachidutswa kakang'ono. Komabe, ngati tsamba loyalikiza likusowa chidutswa cha DNA, kungodzoza kokha kumachitika. M'zosangalatsa zabwino kwambiri, zidutswa zimatha kulerana wina ndi mnzake, koma zokolola zidzakhala zochepa komanso zazing'ono.
. Pa gawo lawotentha, priers idzasungunuka ku matrix DNA ndipo sizingafanane ngakhale pang'ono.
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Vuto lina lomwe limachitika kawirikawiri m'madzi ozindikira ndi ocheperako kuposa momwe zidalili zoperekera ma acid a ma acid poyerekeza ndi chowonjezera cha pchenorofer. Nthawi zambiri, izi zitha kuphatikizidwa ndi zolakwika zabodza. Nthawi yochuluka ikhoza kupulumutsidwa ndi kuwira zinyalala kapena chimbudzi cha ma cell, koma njirayi nthawi zambiri imapangitsa chidwi chotsika cha pcr chifukwa chosakwanira kwa acid.

Zoletsa za polymetrase ntchito mu Kukula

Mwambiri, zoletsa zoletsa zimagwiritsidwa ntchito ngati lingaliro lofotokoza zinthu zonse zomwe zimabweretsa zotsatira za PCR. Mu malingaliro okhazikika biochemical, zoletsa zimangokhala pantchito ya enzyme, ie, imachepetsa kapena zimalepheretsa kutembenuka kwa DNA polymet kapena molt dna polymease).
Zophatikizidwa mu zitsanzo kapena zopindika zosiyanasiyana ndikupanga zomwe zili ndi ma ntchentche kapena kusamalira matope (mwachitsanzo, adatsitsa kapena kuwongolera PCR zotsatira zoyipa za PCR.
Komabe, zokambirana zambiri pakati pa zigawo zikuluzikulu ndi zomwe tikufuna zida zomwe zili ndi zida zimapangidwanso ngati 'PCR inhibitors'. Kukhulupirika kwa khungu kumasokonezedwa ndi kudzipatula komanso ma acidic acid amamasulidwa, kuyanjana pakati pa zitsanzo ndi njira yake yoyatsirana ndi gawo lake loyandikana ndi gawo lolimba lingachitike. Mwachitsanzo, 'Scavenger' imatha kukhala yolumikizirana pang'ono kapena kawirikawiri kudzera pakusintha kosagwirizana ndi kusokoneza kuchuluka kwa ziwerengero zomwe zimayambitsa PCR zomwe zimachitika pambuyo pake.
Mwambiri, pcr zoletsa zimapezeka m'madzi ambiri amayesedwa (ma urea mu mkodzo, hemoglobin, zonunkhira, zitsulo, zitsulo zolemera)

Zowonjezera za PCR zofala zimatha kupezeka mu mabakiteriya ndi ma cell a Eukaryotic, osakhala a DNA, macromolecles a minofu ndi zida zamalonda monga magolovesi ndi pulasitiki. Kudziyeretsa ma acid a nuclec mu nthawi kapena mutachotsa njira yomwe mumakonda kuchotsa PCR zoletsa.
Masiku ano, zida zosiyanasiyana zokhazokha zimatha kusintha ma protocol, koma 100% amachira ndi / kapena kuyeretsa mipherezero sikunakwaniritsidwe. Zolepheretsa zolepheretsa zitha kupezekabe mu asidi woyeretsedwa wa nuclevic kapena mwina adayamba kugwira ntchito. Njira zosiyanasiyana zilipo kuti muchepetse zoletsa. Kusankha kwa polymerase yoyenera kumatha kukhala ndi vuto lalikulu pa ntchito yoletsa. Njira zina zotsimikiziridwa kuti muchepetse zoletsa PCR ikuwonjezera chidwi chowonjezera kapena kugwiritsa ntchito zowonjezera monga BSA.
Zoletsa za pcr zimawonetsedwa ndi kugwiritsa ntchito njira yamkati yowongolera (IPC).
Chisamaliro chimayenera kuchotsedwa kuchotsa ma reagents onse ndi zothetsera zida, monga Ethanol, altcanol ndi phespanol, amasungunuka bwino. Kutengera ndi nkhawa zawo, atha kuyambitsa kapena kuyika pcr.