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PCR tekinoroji yakahwina Mubairo weNobel

Muna 1993, musayendisiti wekuAmerica Mulis akawana Mubairo weNobel muChemistry, uye kubudirira kwake kwaive kugadzirwa kwePCR tekinoroji.Mashiripiti ePCR tekinoroji ari mune anotevera maitiro: Chekutanga, huwandu hweDNA hunofanirwa kukwidziridzwa hudiki zvakanyanya, uye ne theoretically imwe molecule inogona kushandiswa kukwirisa;chechipiri, iyo amplification kunyatsoshanda kwakakwira, uye huwandu hwechinongedzo gene hwakanyanya.Amplification, anopfuura mamirioni gumi nguva mumaawa mashoma.Ikozvino PCR chiridzwa chave kushandiswa zvakanyanya mukutsvagisa hupenyu hwesainzi uye zvimwe zvakawanda zvinhu.

Mhando dzakasiyana uye vagadziri veanopisa cyclers vanogona kuratidza mashandiro akasiyana uye kudzokorora.Iyi misiyano inokanganisa kwete chete kugona kwePCR asiwo iko kurongeka uye kuenderana kwedata rakawanikwa.Kunzwisisa PCR muchina maficha kunogona kutibatsira kuwedzera kubudirira kwezviyedzo zvedu.

Heating module

Iko kurongeka kwekupisa kweiyo thermal cycler kunogona kuita sarudzo yekubudirira kana kutadza kwePCR.Zvakanaka-ku-tsime tembiricha kuenderana pane yekudziya block kwakakoshawo kuti uwane yakavimbika uye inogonekazve PCR mhedzisiro.

Imwe nzira yekuve nechokwadi chekudziya kwekushisa ndeyekuedza kazhinji uchishandisa tembiricha yekuongorora kits uye kugadzirisa zvakare sezvinodiwa nenyanzvi yakadzidziswa.Tembiricha yekuongorora bvunzo inowanzo shandiswa ku:

Zvakanaka-ku-zvakanaka kunyatsoenderana nekuseta tembiricha mune isothermal mode

Zvakanaka-ku-tsime kunyatsoenderana nekuseta tembiricha mushure mekushandura tembiricha

Heat Lid Temperature Kururama

Primer Annealing Temperature Control

Gradient tembiricha yekudzora ibasa rePCR chiridzwa chinofambisa optimization yeprimer annealing muPCR.Chinangwa chekumisikidza gradient ndechekuwana tembiricha dzakasiyana pakati pemamodules, uye ne ≥2 ° C tembiricha inokwira nekudonha pakati pekoramu yega yega, tembiricha dzakasiyana dzinogona kuedzwa panguva imwe chete kuti uwane iyo yakakwana primer annealing tembiricha.Theoretically, yechokwadi gradient inowana mutsara tembiricha pakati pemamodule.

Nekudaro, akajairwa gradient thermal cyclers anowanzo shandisa imwechete yekupisa uye yekudzora tembiricha kuburikidza maviri ekudziya uye kutonhora zvinhu zviri kumagumo ese, kazhinji zvichikonzera zvinotevera zvinogumira:

Matembiricha maviri chete anogona kusetwa: tembiricha yakakwira uye yakaderera yeprimer annealing yakaiswa pamagumo ese eiyo thermal module, uye kurongeka chaiko kwemamwe tembiricha hakugone kuwanikwa pakati pemodul.

Nekuda kwekuchinjana kwekupisa pakati pemakoramu akasiyana, tembiricha pakati penzvimbo dzakasiyana pane moduru inowanzotevera sigmoidal curve pane yechokwadi mutsara gradient.

Sample tembiricha

Iko kugona kweiyo thermal cycler kudzora tembiricha yekuenzanisira kwakakosha kune chokwadi chemhedzisiro yePCR.Instrument-chaiyo paramita senge ramp rates, nguva dzekubata, uye algorithms zvakakosha pakufanotaura tembiricha yemuenzaniso.

Mwero wekudziya nekutonhodza kwemuchina wePCR zvinoreva kuti tembiricha inoshanduka pakati pematanho ePCR anoitika pane imwe nguva yenguva.Sezvo zvinotora imwe nguva yenguva kuti kupisa kuende kubva mumushonga kuenda kumuenzaniso, iyo chaiyo yekupisa uye kutonhora kwemuenzaniso kuchave kunonoka.Naizvozvo, tsananguro yekumhanya kwekuchinja kwekushisa inoda kusiyaniswa uye kunzwisiswa.

Iyo yakanyanya kana peak module ramp rate inomiririra inokurumidza kuchinja tembiricha iyo module inogona kuwana pamusoro penguva diki diki yenguva panguva yerampu.

Avhareji block ramp rate inomiririra chiyero chekuchinja kwekushisa kwenguva yakareba uye ichapa imwe inomiririra chiyero chePCR muchina kumhanya.

Iyo yakanyanya sampuli yekudziya uye kutonhora mwero uye avhareji sampuli yekudziya uye kutonhora mwero inoratidza tembiricha chaiyo inowanikwa nesample.Chiyero chekudziya kwemuenzaniso uye kutonhora chinopa kuenzanisa kwakaringana kwekuita kwePCR muchina uye kukanganisa kwayo pamhedzisiro yePCR.

Paunenge uchiita cycler kutsiva, zvinokurudzirwa kushandisa chiridzwa chine ramp rate chirongwa chinoteedzera yakapfuura modhi yekutsiva nyore uye kushoma kukanganisa paPCR kudzokorora.

Iyo inopisa cycler inofanirwa kugadzirwa kuti iite matanho chete mushure mekunge sampu yasvika patembiricha yakatarwa.Nenzira iyi, iyo nguva iyo sampu inochengeterwa pakushambidzika kweseti ichave yakanyatso chengetedzwa neinopindirana kutenderera mamiriro anodiwa mune yekushanda maitiro.

Thermal cyclers inowanzo shandisa yakaoma mathematical algorithms kuona kuti masampuli anogona kukurumidza kusvika patembiricha yakatarwa zvinoenderana nechirongwa chakafanorongwa.Zvichienderana nehuwandu hwemaitiro ekuita uye kukora kwePCR mapurasitiki anoshandiswa, iyo algorithm inogona kufanotaura tembiricha yesample uye nguva yaichatora kuti isvike tembiricha yakatarwa.Tichivimba neaya maalgorithms, panguva yekudziya kana kutonhora kweiyo thermal cycler, tembiricha yeblock inowanzopfuura kukosha kweseti kuburikidza nemaitiro anonzi thermal block overshoot kana undershoot.Kugadzika kwakadaro kunovimbisa kuti sampuli inosvika pakushisa kweseti nekukurumidza sezvinobvira pasina overshooting kana undershooting pachayo.

Experimental throughput

Zvinhu zvinogona kuwedzera kubuda kweiyo inopisa cycler zvinosanganisira ramp rates, thermal block configurations, uye kubatanidzwa kweautomation mapuratifomu.

Kupisa nekutonhodza mwero we thermal cycler inomiririra kumhanya kwainosvika patembiricha yakatarwa.Iyo inokurumidza kukwira kwekushisa nekudonha, iyo inokurumidza PCR inomhanya, izvo zvinoreva kuti mimwe miedzo inogona kupedzwa munguva yakatarwa.Pamusoro pezvo, zviedzo zvinogona kukwidziridzwa nekushandisa nekukurumidza DNA polymerases.

Dhizaini yeiyo thermal cycler module yakakoshawo kune PCR kuyedza.Semuenzaniso, mamodule anogona kuchinjika anobvumira kuchinjika muhuwandu hwemasampuli pakumhanya.Uye zvakare, kudziyisa mamodule ane ega ega anodzoreka mamodule akanakira kumhanya akasiyana PCR zvirongwa panguva imwe chete pane imwe thermal cycler.

Kune otomatiki yakakwira-kuburikidza PCR, software inodzora iyo pipetting inobata system inofanirwa kurongeka uye inoenderana.Automated systems dzakanakira kuita high-throughput PCR reactions nekuti dzinogona kuitwa nguva dzose nekupindira kwevanhu kushoma, nekudaro kuderedza nguva inodiwa pakuseta kwekuedza uye kuwedzera huwandu hwekuita munguva yakatarwa.

Kuvimbika, Kusimba uye Hunhu Kuvimbiswa kweThermal Cyclers

Pamusoro pekuita uye kugona kwekupinda, muchina wePCR unofanirwawo kushingirira kumwe kushandiswa kunodzokororwa, kushushikana kwezvakatipoteredza, uye mamiriro ekutumira.Vamwe vagadziri vanogona kutaura nezve mashandisiro anoita chiridzwa kuvimbika uye kusimba bvunzo.Iyo inoenderana PCR chiridzwa yekuona inosanganisira:

Kuvimbika: Mechanical rigs anoshandiswa kuita bvunzo dzakadzokororwa pazvinhu zvinogara zvichishandiswa zviridzwa senge thermal lids, control panel / touchscreens, uye tembiricha cycling modules.

Ambient pressure: Dzimba dzezvakatipoteredza dzinogona kushandiswa kutevedzera mamiriro akasiyana ezviyedzo zvenguva dzose, senge tembiricha, humidity.

Kuongorora Kwekutumira: Kuvhundutsa kwakanyanya uye kuyedza kuvhiyiwa kunogona kuitwa zvinoenderana neInternational Safety Shipping Association zviyero kuti ive nechokwadi chekuti chiridzwa chinogona kusvika mumamiriro ekushanda asina kukuvara.

Warranty uye sevhisi yekuchengetedza PCR muchina

Kunyangwe kuvimbika kwakasimba uye kusimba kwekuyedzwa, mabhasikoro anopisa ane nyaya dzehunyanzvi pamusoro pehupenyu hwechiridzwa.Kuti uve nerunyararo rwepfungwa, waranti yemugadziri, sevhisi, uye kugadzirisa kunofanirwa kutariswa kana uchitenga chiridzwa.

Iko kuchinjika kwemasevhisi akadai sepa-saiti/kudzokera-kufekitari kugadzirisa, kure kure yekutarisa masevhisi, uye ekutsiva zviridzwa mukugadzirisa maitiro, nezvimwewo, kuderedza kukanganiswa kwekushanda kwekushanda.

Hurefu hwenguva yewaranti, nguva yekushandura yebasa, kuwanikwa kwerutsigiro rwehunyanzvi, uye hunyanzvi hwevashandi vanotsigira.

Kugoneka kwekumisikidzwa kwechiridzwa, kushanda, kushandira pamwe, uye kuonesa kusangana nerabhoritari uye zvinoenderana nezvinodiwa zvemitemo.Masevhisi ekuchengetedza akadai sekutarisa tembiricha, kuyedzwa, uye calibration anowanikwa kuti ave nechokwadi chekuti chiridzwa chinoshanda nemazvo nemaparamendi anoenderana.

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