Injin PCR|Shin da gaske kuna fahimta?

Fasahar PCR wadda ta lashe kyautar Nobel

A shekarar 1993, masanin kimiyar Amurka Mulis ya samu kyautar Nobel a fannin ilmin sinadarai, kuma nasarar da ya samu ita ce kirkiro fasahar PCR.Sihiri na fasaha na PCR yana cikin halaye masu zuwa: Na farko, adadin DNA ɗin da za a ƙarawa yana da ƙanƙanta, kuma a ka'ida ana iya amfani da kwayoyin halitta ɗaya don haɓakawa;na biyu, ingancin haɓakawa yana da yawa, kuma adadin ƙwayar da aka yi niyya yana da yawa.Ƙarawa, fiye da sau miliyan 10 a cikin 'yan sa'o'i.Yanzu an yi amfani da kayan aikin PCR sosai a cikin binciken kimiyyar rayuwa da sauran fannoni da yawa.

Samfura daban-daban da masana'antun masu kekunan zafin jiki na iya nuna ayyuka daban-daban da maimaitawa.Waɗannan bambance-bambance suna shafar ba kawai ingancin PCR ba har ma da daidaito da daidaiton bayanan da aka samu.Fahimtar fasalin injin PCR zai iya taimaka mana haɓaka nasarar gwajin mu.

Tsarin dumama

Daidaiton zafin jiki na thermal cycler na iya zama yanke hukunci don nasara ko gazawar PCR.Matsakaicin yanayin zafi mai kyau zuwa rijiyar akan toshewar dumama shima yana da mahimmanci don samun ingantaccen sakamako na PCR wanda za'a iya maimaitawa.

Hanya ɗaya don tabbatar da daidaiton zafin jiki shine a gwada akai-akai ta amfani da na'urorin tabbatar da zafin jiki da sake daidaitawa kamar yadda ƙwararren ƙwararren ƙwararren ya buƙata.Ana amfani da gwaje-gwaje na tantance zafin jiki don:

Daidaito-zuwa rijiya dangane da saita zafin jiki a yanayin isothermal

Daidaito-zuwa rijiya dangane da saita zafin jiki bayan canjin yanayin zafi

Daidaiton Rufe Zafi

Sarrafa zafin jiki na Farko

Sarrafa zafin jiki na gradient aiki ne na kayan aikin PCR wanda ke sauƙaƙa inganta haɓakar haɓakar firam a cikin PCR.Manufar saitin gradient shine don cimma yanayin zafi daban-daban tsakanin kayayyaki, kuma ta ≥2°C yanayin zafi da faɗuwa tsakanin kowane ginshiƙi, ana iya gwada yanayin zafi daban-daban lokaci guda don samun mafi kyawun zafin jiki na share fage.A ka'ida, gradient na gaskiya yana samun zafin jiki na madaidaiciya tsakanin kayayyaki.

Koyaya, masu hawan keke na al'ada na al'ada suna amfani da toshewar thermal guda ɗaya da zafin jiki ta hanyar dumama abubuwa biyu masu sanyaya da ke kusa da ƙarshen biyu, galibi suna haifar da iyakoki masu zuwa:

Za'a iya saita yanayin zafi guda biyu kawai: babban yanayin zafi da ƙananan zafin jiki don annealing na farko an saita su a ƙarshen tsarin thermal, kuma ba za a iya samun daidaitaccen yanayin sauran yanayin ba tsakanin modul.

Saboda musayar zafi tsakanin ginshiƙai daban-daban, zafin jiki tsakanin yankuna daban-daban akan tsarin yana da yuwuwar bin madaidaicin sigmoidal maimakon madaidaiciyar madaidaiciyar madaidaiciya.

Samfurin zafin jiki

Ƙarfin mai zazzagewa na thermal don sarrafa zafin samfurin yana da matukar mahimmanci don daidaiton sakamakon PCR.Ƙayyadaddun ƙayyadaddun kayan aiki kamar ƙimar ramp, lokutan riƙewa, da algorithms suna da mahimmanci don tsinkayar zafin samfurin.

Adadin dumama da sanyaya na'urar PCR yana nufin cewa yanayin zafi yana canzawa tsakanin matakan PCR da ke faruwa a kan wani ɗan lokaci.Tun da yake yana ɗaukar wani lokaci don zafi don canjawa daga samfurin zuwa samfurin, ainihin dumama da sanyi na samfurin zai kasance a hankali.Don haka, ma'anar saurin canjin zafin jiki yana buƙatar rarrabewa da fahimta.

Matsakaicin matsakaicin matsakaicin ko mafi girma na ƙirar ƙirar yana wakiltar canjin zafin jiki mafi sauri wanda tsarin zai iya cimma kan ɗan kankanin lokaci yayin hawan.

Matsakaicin adadin toshewa yana wakiltar ƙimar canjin zafin jiki na tsawon lokaci kuma zai samar da ƙarin ma'aunin wakilci na saurin injin PCR.

Matsakaicin adadin dumama da sanyi da matsakaicin samfurin dumama da sanyi suna nuna ainihin zafin jiki da samfurin ya samu.Adadin samfurin dumama da sanyaya zai samar da ingantaccen kwatancen aikin injin PCR da yuwuwar tasirinsa akan sakamakon PCR.

Lokacin yin maye gurbin mai keke, ana ba da shawarar yin amfani da kayan aiki tare da shirin ƙimar ramp wanda ke kwatanta yanayin da ya gabata don sauƙin sauyawa da ƙaramin tasiri akan maimaitawar PCR.

Ya kamata a ƙera na'urar zazzagewar zafi zuwa matakan lokaci kawai bayan samfurin ya kai yanayin zafin da aka saita.Ta wannan hanyar, lokacin da aka kiyaye samfurin a yanayin zafin da aka saita zai kasance mafi dacewa tare da yanayin sake zagayowar da ake buƙata a cikin tsarin aiki.

Masu kekuna masu zafi sukan yi amfani da hadadden algorithms na lissafi don tabbatar da cewa samfurori na iya isa ga yanayin zafin jiki da sauri bisa ga tsarin saiti.Dangane da girman tsarin amsawa da kauri na robobin PCR da aka yi amfani da su, algorithm na iya yin hasashen yanayin zafin samfurin da lokacin da zai ɗauka don isa ga yanayin da aka saita.Dogaro da waɗannan algorithms, yayin aikin dumama ko sanyaya na'urar zazzagewar, yawan zafin jiki na toshe zai wuce ƙimar da aka saita ta hanyar tsari da ake kira thermal block overshoot ko undershoot.Irin wannan saitin yana tabbatar da cewa samfurin ya kai ga yanayin da aka saita da sauri ba tare da wuce gona da iri ba ko kuma a kasa harbi kanta.

Fitar da gwaji

Abubuwan da za su iya ƙara yawan kayan aikin mai zazzagewar zafin jiki sun haɗa da ƙimar ramp, ƙayyadaddun toshewar zafi, da haɗin dandamali na sarrafa kansa.

Matsakaicin dumama da sanyaya na mai zazzagewar zafi yana wakiltar saurin da ya kai yanayin da aka saita.Da sauri tashin zafin jiki da faɗuwa, da sauri PCR zai gudana, wanda ke nufin za a iya ƙara ƙarin gwaje-gwaje a cikin wani ɗan lokaci.Bugu da ƙari, ana iya ƙara gwaje-gwaje ta hanyar amfani da polymerases na DNA da sauri.

Zane na thermal cycler module shima yana da mahimmanci don gwaje-gwajen PCR.Misali, samfuran da za'a iya maye gurbinsu suna ba da damar sassauci a cikin adadin samfuran kowane gudu.Bugu da kari, na'urorin dumama tare da na'urori masu sarrafawa daban-daban sun dace don gudanar da shirye-shiryen PCR daban-daban a lokaci guda akan na'ura mai zafi guda ɗaya.

Don PCR mai girma mai sarrafa kansa, software ɗin da ke sarrafa tsarin sarrafa bututu ya kamata ya zama mai tsari kuma ya dace.Tsarin sarrafa kansa yana da kyau don aiwatar da halayen PCR mai girma saboda ana iya aiwatar da su gabaɗaya tare da ɗan ƙaramin saƙon ɗan adam, ta haka rage lokacin da ake buƙata don saitin gwaji na hannu da haɓaka adadin halayen a cikin ɗan lokaci.

Dogara, Dorewa da Tabbacin Ingancin Na'urorin Zazzabi

Baya ga iyawar aiki da kayan aiki, injin PCR ya kamata kuma ya iya jure wasu maimaita amfani, matsalolin muhalli, da yanayin jigilar kaya.Wasu masana'antun na iya bayar da rahoto kan yadda kayan aikin ke yin amintacce da gwajin dorewa.Daidaitaccen gano kayan aikin PCR ya haɗa da:

Dogaro: Ana amfani da injin injina don yin gwaje-gwaje akai-akai akan kayan aikin da aka saba amfani da su akai-akai kamar murfi mai zafi, bangarorin sarrafawa/allon taɓawa, da na'urorin hawan keke na zafin jiki.

Matsin yanayi: Za a iya amfani da ɗakunan muhalli don daidaita yanayi daban-daban na gwaje-gwaje na yau da kullum, kamar zafin jiki, zafi.

Gwajin jigilar kaya: Za a iya yin gwajin girgiza da girgizawa bisa ga ƙa'idodin Ƙungiyar Tsaro ta Duniya don tabbatar da kayan aikin na iya isa cikin yanayin aiki mara lahani.

Garanti da sabis don kiyaye injin PCR

Duk da tsayayyen aminci da gwajin dorewa, babu makawa masu hawan zafi suna da al'amuran fasaha akan rayuwar kayan aikin.Don kwanciyar hankali, garantin masana'anta, sabis, da kiyayewa yakamata a yi la'akari da lokacin siyan kayan aiki.

Sauƙaƙe na ayyuka kamar a kan-site / koma-zuwa masana'antu kiyayewa, m saka idanu ayyuka, da kuma maye kayan aiki a cikin tsarin kulawa, da dai sauransu, don rage tasiri a kan ingancin aiki.

Tsawon lokacin garanti, lokacin juyawa na sabis, samun damar tallafin fasaha, da ƙwarewar ƙwararrun ma'aikatan tallafi.

Yiwuwar shigar kayan aiki, aiki, haɗin gwiwa, da tabbatarwa don saduwa da dakin gwaje-gwaje da buƙatun tsari masu alaƙa.Ana samun sabis na kulawa kamar tabbatar da zafin jiki, gwaji, da daidaitawa don tabbatar da cewa kayan aikin yana yin aiki yadda ya kamata tare da madaidaitan sigogi.

Samfura masu dangantaka: