Kugenzura imikorere ya primers na probe mubyiciro byambere bya reagent ya PCR no kumenya uburyo bwiza bwo kubyitwaramo nibisabwa kugirango habeho iterambere ryibigeragezo bisanzwe.
Nigute dukeneye kwemeza primer probe mugihe cyambere?
Ibipimo nyamukuru ni ibyingenzi, amplification curve, ct agaciro, amplification efficient, sample-concentration sample detection, CV, nibindi.
Ibyingenzi
Ibyingenzi ni umurongo utambitse muri PCR amplification curve.Mubice bike byambere bya PCR amplification reaction, ibimenyetso bya fluorescence ntabwo bihinduka cyane kandi bikora umurongo ugororotse.Uyu murongo ugororotse niwo shingiro.
Mugihe cyo kwerekana PCR primer probe, witondere niba ibyingenzi ari urwego.Ubuziranenge bwibisobanuro bya primer probe bizagira ingaruka kumurongo, nko gutera umurongo kuzamuka cyangwa kugwa.Ibyingenzi nabyo ni ibimenyetso byerekana neza.
Amplification Curve
Ikindi kimenyetso cyerekana ni imiterere ya amplification curve.Nibyiza kugira umurongo S ufite umurongo kugirango wirinde amplification ya kabiri cyangwa indi ntera idasanzwe idasanzwe.
Ct Agaciro
Umubare wizunguruko uhuye nu guhinduranya kuva kumurongo wibanze kugera kumikurire yerekana ni Ct agaciro.
Kuri sample imwe, primer probe zitandukanye zitanga ibisubizo bitandukanye byo kwongera umurongo, kandi agaciro ka Ct kajyanye ningaruka ziterwa no kongera imbaraga hamwe nimpamyabumenyi.Mubyigisho, ntoya Ct agaciro ka primer probe duhitamo, nibyiza.
Gukora neza
Bumwe mu buryo bwizewe kandi butajegajega bwo gusuzuma imikorere ya PCR ikora neza ni umurongo usanzwe, nawo uzwi n'abashakashatsi.Uburyo bukubiyemo gukora urukurikirane rwintangarugero kugirango ugenzure umubare ugereranije wintego.Izi ngero zisanzwe zikorwa nuruhererekane rwibisubizo byibisubizo byibisubizo, bikunze gukoreshwa ni inshuro 10.Ukoresheje urukurikirane rwicyitegererezo, ukoresheje progaramu isanzwe ya qPCR kugirango wongere kugirango ubone agaciro ka Cq, hanyuma ushushanye gushushanya umurongo usanzwe ukurikije ubunini bwa buri sample hamwe nagaciro ka Cq bihuye kugirango ubone umurongo ugereranije Cq = -klgX0 + b, hamwe no kongera imbaraga E = 10 (-1 / k) -1.Iyo ukoresheje qPCR mu gusesengura ingano, imikorere ya amplification isabwa kuba iri hagati ya 90% -110% (3.6> k> 3.1).
Icyitegererezo Cyibanze Cyicyitegererezo
Iyo icyitegererezo cyibanze kiri hasi, igipimo cyo gutahura primer probe zitandukanye ziratandukanye.Duhitamo ibyitegererezo 20 byo hasi kugirango twigane, kandi sisitemu ya primer-probe hamwe nigipimo kinini cyo gutahura nibyiza.
Coefficient de Variation (CV)
Ingero 10 zibiri zirashobora gutahurwa hamwe na primer probe zitandukanye ukurikije umurongo wa reagent kugirango tumenye aside nucleic aside.
Umubare wuzuye:
Ukuri
Ukuri mubice bimwe bigomba guhura: coefficente yo gutandukana (CV,%) yagaciro ka logarithmic yibipimo byibizamini ni ≤5%.Iyo icyitegererezo cyibanze kiri hasi, coefficente yo guhinduka (CV,%) ya logarithm yibitekerezo byo kumenya ni ≤10%
Imiterere ya reagent:
Ukuri
Ukuri mubice bimwe bigomba guhura:
(1) Coefficient yo gutandukanya agaciro ka Ct (CV,%) ≤5%
Icyitegererezo kimwe cyageragejwe mugihe cyinshuro 10, kandi ibisubizo byikizamini bigomba kuba bihamye
Igihe cyo kohereza: Nzeri-18-2021
