1: Ku beddel saadka tijaabada wakhtiga

Deji (NTC) xakamaynta taban oo ku celi marar badan.Marka la ogaado inay jirto wasakhaynta alaabta PCR ee shaybaarka, beddel dhammaan sahayda tijaabada wakhtiga.Sida: dib u milo oo diyaari furayaasha, dib u nadiifi caarada pipette, tuubada EP, ddH2O, iwm., ku beddel pipette cusub, oo si ku meel gaar ah u amaahdo shaybaarro kale si aad u sameyso tijaabooyinka PCR.Shaybaadhka wasakhaysan waa in la neefsadaa oo lagu shubaa fallaadhaha ultraviolet ilaa inta wasakhdaynta PCR laga saarayo ka hor inta aan la sii wadin tijaabada.

2: Kordhi wakhtiga soo-gaadhista UV

Waxaa xusid mudan in si looga saaro wasakheynta DNA-da, shucaaca ultraviolet ee caadiga ah waa in lagu kordhiyo 2 saacadood intii caadiga ahayd.Si kastaba ha ahaatee, saameynta shucaaca UV ee ka saarista jajabyada yaryar (ka hooseeya 200bp) ee wasakheynta DNA wali ma fiicna.

Mawjada Ultraviolet (nm) guud ahaan waa 254/300nm, waxayna ku filan tahay in la shido 30 daqiiqo.Waa in la ogaadaa in marka la dooranayo UV si loo baabi'iyo wasakhda alaabta PCR ee haraaga ah, dhererka alaabta PCR iyo qaybinta saldhigyada isku xigxiga alaabta waa in la tixgeliyaa.Shucaaca UV ayaa kaliya waxtar u leh jajabyada dhaadheer ee ka sarreeya 500 bp, waxayna saameyn yar ku leedahay jajabyada gaaban.

Inta lagu jiro shucaaca UV, saldhigyada pyrimidine ee alaabta PCR waxay sameyn doonaan dimers.Dimers-yadani waxay joojin karaan kordhinta, laakiin dhammaan pyrimidine ee silsiladda DNA ma samayn karaan dimers, iyo shucaaca UV ayaa sidoo kale jebin kara dimers..Heerka samaynta dimer waxay ku xidhan tahay hirarka UV, nooca pyrimidine dimer iyo taxanaha nucleotides ee ku dheggan goobta dimer.Sidaa darteed, haddii jajabyada la xoojiyay ee PCR ay yar yihiin, waxaa lagu talinayaa in la isticmaalo nidaamka wasakheynta alaabta UNG anti-PCR.

3:Wasakhowga DNA-da ee sida caadiga ah loo isticmaalo

Aerosols si fudud ayaa loo soo saaraa marka tuubooyinka lagu daro, taas oo ay adag tahay in laga fogaado, si dhakhso ah ayey u degi doontaa.Sidaa darteed, shaki la'aan waa doorasho wanaagsan in si joogto ah loo isticmaalo qashinka wasakhowga DNA-ga gaarka ah si looga hortago faafitaanka wasakhowga DNA-da.

4:Isticmaal habka ka hortagga wasakhowga ee UNG

Ka dib marka wasakhdaynta alaabta PCR meesha laga saaro, shaybaadhka baadhista waxa uu isticmaali karaa habka wasakhaynta alaabta UNG anti-PCR si looga hortago wasakhaynta alaabta PCR.Shaybaadhyada guud, waxaad samayn kartaa qaybo tijaabo ah oo fudud, si adag uga sooc aagga alaabta PCR ee meelaha kale, dejiso xeerar iyo qawaaniin shaybaadhka qaarkood, oo aad qabato tababar ku habboon si looga hortago inay dhacdo wasakhowga alaabta PCR.

Talooyin: Samaynta shaybaadhka PCR ee macquulka ah, ilaalinta deegaanka PCR wanaagsan, samaynta habraacyada hawlgalka PCR caadiga ah, iyo kobcinta wacyiga adag ee hawl wadeenada ayaa ah furayaasha ka hortagga ama yaraynta wasakhowga tijaabooyinka PCR.