Polymerase chain reactions (PCR) ndeimwe yenzira dzinozivikanwa zvikuru dzinoshandiswa mumarabhoritari esainzi yehupenyu.
Iwo maPCR mahwendefa anogadzirwa kubva ekutanga-kirasi mapurasitiki ekugadzirisa zvakanakisa uye kuongororwa kwemasampuli kana mhedzisiro inounganidzwa.
Vane madziro akatetepa uye ane homogenous kuti ape chaiyo thermal transfer.
Mukugadzirira mashandisirwo enguva chaiyo, chikamu cheminiti cheDNA kana RNA chakavharirwa uye chakachengetwa mumapureti ePCR.
Iwo PCR mahwendefa anoshanda zvakanyanya mukupisa chisimbiso uye anodzivirira kuyerera kwekupisa zvakare.
Nekudaro, seanoshanda uye akavimbika maPCR mahwendefa ari, kukanganisa uye kusarongeka kunomira nyore paunenge uchigadzira samples.
Naizvozvo, kana iwe uchida kuwana yakanaka uye yepamusoro mhandoPCR mahwendefa.Zvakanakira kubata akavimbika PCR ndiro inogadzira. Neizvi une chokwadi chekuwana dhiri yakanakisa.
Heano mamwe matanho ekutevera kudzivirira kusvibiswa kwemareagents kana samples uye kudzivirira kusarongeka kubva mukukwira mumhedzisiro.
Kusunga Matunhu
Zvisizvo zvakanaka kana zvisizvo zvinoitika nekuda kwekuvapo kwetsvina, izvo zvinoita kuti usave nechokwadi nemhedzisiro.
Kusachena uye kusvibiswa kunoitika nenzira dzakasiyana-siyana senge isina hukama DNA kana makemikari ekuwedzera izvo zvinopedzisira zvadzikisira kugona uye kushanda kwekuita.
Kune nzira dzakawanda dzekudzikisa zvakanyanya mwero wekusvibiswa kwePCR plate.
Kushandisa sterilized filter tips ndiyo imwe nzira inobatsira yekudzivirira tsvina kuti isapinda mumasampuli ako kuburikidza nemapipette.
Shandisa seti yakachena yemidziyo, inosanganisira pipettes uye racks, chete kushandiswa kwePCR. Izvi zvinovimbisa kutamiswa kusingaite kwetsvina kana zvinosvibisa zvakatenderedza murabhoritari.
Shandisa mableaches, ethanol pamapipettes, racks nemabhenji kupukuta zvinosvibisa.
Govera nzvimbo yakachengeterwa yako yese PCR maitiro kuti uwedzere kudzikisira kusvibiswa kwechikamu.
Shandisa magirovhosi akachena pamatanho ese uye woatsiva kazhinji.
PCR mahwendefa
Ongorora Kudzika uye Kuchena kweTemberi.
Kuchena kwebhenji uye midziyo inoshandiswa pakuongorora samples nePCR inofanira kuchengetedzwa. Izvo zvakakosha kuti usimbise dhigirii rekuchena kwemasampuli usati waongorora uye kugadzirisa.
Kazhinji, vanoongorora vanotarisisa kutariswa uye kuchena mwero weiyo DNA samples.
Edza chiyero chekunwa kwe260nm/280nm haifanire kunge iri pasi pe1.8. Nepo hunotevera wavelength pakati 230nm uye 320nm inoshandiswa kuona tsvina.
Pane imwe nguva, chaotropic munyu uye mamwe organic komisheni anoonekwa pa230nm yekunyudza mwero. Nepo turbidity muDNA samples inowonekwawo nekusimbiswa pamwero wekutora 320nm.
Dzivisa kuwandisa PCR Plates nechigadzirwa
Zvakawanda sekudiwa kumhanyisa zvigadzirwa zvakawanda panguva imwe chete, zvinokonzeresa kusvibiswa kwePCR mahwendefa.
Kuremedza mahwendefa ePCR nemarara akasiyana ezvigadzirwa uye zvinoita kuti zvinyanye kunetsa kuwana masampuli.
Chengetedza Zvinyorwa zveAliquot PCR Reagents
Kuenderera mberi kwechando/kunyunguduka kutenderera uye kugara uchishandiswa kwealiquot kunogona kukanganisa PCR reagents, ma enzymes uye DNTP's kuburikidza nekugadzirisa zvakare.
Gara uchiedza kutarisa mwero wealiquot inoshandiswa paunenge uchigadzira masampuli kuti aongororwe.
Inofarirwa LIMS inonyanya kukodzera kudzora hesitori uye huwandu hwema reagents uye samples dzakaoma kana kunyungudika.
Sarudza Iyo Yakanakisa Annealing Temperature.
Kutora uye kushandisa tembiricha isiriyo yekubatisa ndiyo imwe nzira PCR mhedzisiro ine kukanganisa.
Dzimwe nguva, mhinduro yacho haiendi sezvakarongwa. Inoshuviwa kudzikisa tembiricha ye annealing kuitira kufambisa kuita kwakabudirira.
Nekudaro, kudzikisa tembiricha kunowedzera mikana yenhema positives uye primer dimers kutaridzika.
Izvo zvakakosha kusimbisa kuongororwa kwekunyunguduka curve kana uchishandisa PCR mahwendefa sezvo chiri chiratidzo chakanaka chekusarudza tembiricha chaiyo yekunyungudika.
Primer dhizaini software inobatsira nekugadzira, kupihwa kweiyo annealing tembiricha ine zvakananga inoderedza kukanganisa muPCR mahwendefa.
Mukuda Yepamusoro -Unhu PCR ndiro?
Kana iwe wanga uchifunga kuti ungawane kupi mugadziri akavimbika wePCR mahwendefa. Usatsvagezve nekuti wasvika panzvimbo chaipo.
Nomutsatinya pano kuti ubate nesuzvemhando yepamusoro zvigadzirwa uye sevhisi pamutengo usingaputse bhangi.
Nguva yekutumira: Oct-30-2021