Isolating DNA kubva E. Coli hunhu hwakakosha mumanyorerwo ebiology. Chinyorwa ichi chinokufambisa iwe kuburikidza nemaitiro ese, kupa matanho akatsanangurwa uye tsananguro, ichiva nechokwadi chekuti unonzwisisa mese sainzi uye zvinhu zvinoshanda zvehunhu. Kunyangwe iwe uri muongorori ane ruzivo kana mutsva weiyo lab, iyi nhungamiro ichave chinhu chakakosha chivakwa.
Kugadzirira kwekumiswa kweCell
● Kuunganidzwa kweE. Coli Masero
Nhanho yekutanga mukuparadzanisa DNA kubva kuE. Coli inosanganisira kuunganidza masero ehutachiona. Izvi zvinowanzoda kukura e. coli mune yakakodzera mvura svikiro kusvika yasvika padanho rekuzara ratidziro. Nguva yakakosha nekuti masero ari muchikamu ichi anonyanya kushanda uye ari nyore kufambisa, izvo zvichaguma nekukohwa kwakanyanya.
● Masero anotaurisa mune yakakodzera buffer
Masero akaunganidzwa anobva aunganidzwa mune yakakodzera buffer. Chisarudzo chakajairika ndeye tris - Edta (te) buffer, iyo inobatsira kuchengetedza kutendeka kweDNA panguva yekutsamwa. Iyo buffer inoshandisa zviyero zvakawanda: Inoita kuti pH, inodziya zvikamu zvekuvanza izvo zvinogona kuchinja DNA, uye inopa yakakwana Ionic nharaunda yekuita enzymatic maitiro.
Centrifugation kuenda kumasero ellet
● Paramende yeCentrifugation (Kumhanya uye Nguva)
Mushure mekusunungura masero, kumiswa kunoiswa kuCentrifugation kuenda kuPellet the masero. Centrifugation kumhanya uye nguva yakanyanya kuoma parameter. Kazhinji, centrifugation inoitwa panguva dzinenge 4,000 - 6,000 g for 10 - 15 maminitsi pa 4 ° C. Izvi zvinovimbisa kuti masero anoumba pellet yakasimba pasi pe centrifige tube.
● Kukosha kwekucheka kwakakodzera
Kuchekera kwakakodzera kwakakosha kupatsanura masero kubva kune yekukura kwepakati uye zvimwe zvinhu zvinonyadzisira. Tsime - Yakaumbwa Pellet inoita kuti matanho anotevera ari nyore uye anoshanda zvakanyanya, nekuita kurasikirwa kushoma kwemasero uye, saka, Maximum Scored DNA will.
Kubviswa kwe supernatant
● Matekota ekubvisa supernatant kubviswa
Kana masero akamboputirwa, iyo supernatant (iyo mvura pamusoro peiyo pellet) inofanira kunyatso bviswa isina kuvhiringidza iyo pellet. Izvi zvinowanzoitwa uchishandisa micropipette. Zvakakosha kuti uite nhanho iyi zvine hunyanzvi kuti usarasikirwa nemasero chero.
● Kuvhiringidza kurasikirwa kushoma kwefellet
Kuverengera kushoma kurasikirwa kweiyo cell pellet inosanganisira kuchenjera pipetting uye, kana zvichikosha, akawanda mataundi ezasi okutanga uye kuregedza kureba. Chinangwa ndechekuchengeta masero mazhinji sezvinobvira muPellet yeellet yeexmope yekuwisira.
Kuwedzera kweNuclei Lysis Solis Solis Soliss
● Zvikamu zveNuclei Lysis Solis Soliss
Iyo Nuclei Lyiss Solis Solis Solis Solis Soliss ine dhizaini (kunge sds), buffer (senge tris - hcl), uye yekunyepedzera (seyakarukwa (seEdta). Kudonhedza kunokanganisa iyo mendang metrane uye enveloar yeNyukireya, kusunungura iyo celluly zvirimo, kusanganisira DNA, mune mhinduro.
● Basa rekuputsa masero cell
Iyo Nuclei Lyiss Solis Solis Sosi Haisi chete Memberane yeSero asiwo kuyananisa mapuroteni uye maLipids kuputsa madziro emusero uye envelear envelopu yekuburitsa dhirowa mumhinduro.
RegereSpension yemasero
● Nyorova pipetting kuti udzivise kufura
Kamwe chete iyo nyule lylsis mhinduro inowedzerwa, masero anofanirwa kutangwazve zvinyoronyoro kudzivirira kugura. Kuveura kunogona kupwanya iyo zvidiki zvidiki, izvo zvinogona kuve zvinonetsa zvekusimbisa kwekushandisa izvo zvinoda kukwirira - Molecular - Weightur.
● Kuona kuzorora kwakazara
Kupedzisa kuzorora kunovimbisa kuti masero ese ari kufanana neyakafanana, ekukurisa DNA kupora. Izvi zvinogona kuwanikwa neyenyoro pipetting kana vortexing iyo mhinduro pane yakaderera kumhanya.
Incubation kune lyse masero
● Tembiricha yekuisa yeChinyorwa
Masero ezvekutanga akavezwa pane tembiricha chaiyo kuti ive nechokwadi chekupedzisira lyis. Izvi zvinowanzoitwa pa37 ° C kusvika 55 ° C. Iyo tembiricha chaiyo uye nguva inokwanisa kusiyana zvichienderana ne protocol uye zvakananga zvinodiwa zveDNA yekuzviparadzanisa kit yekushandiswa.
● Nguva inodiwa kuti inyararirel
Nguva yakajairika yenguva yekukwana iri pakati pemaminetsi makumi maviri kusvika makumi matatu, asi inogona kuchinjwa zvichienderana nekubudirira kwesero kucherechedzwa. Kwenguva refu incubation inogona kunge iri madikanwa kuti lyis izere asi inofanira kuve yakaringana kubva kune njodzi yeDNA DNAgradation.
Kutonhora kumhepo tembiricha
● Kukosha kwekutonhora zvishoma nezvishoma
Mushure mekunge mugumo, iyo lysate inodonhedza zvishoma nezvishoma kumhepo tembiricha. Zvishoma nezvishoma kutonhora kunobatsira mukudzikisira iyo DNA uye kuderedza njodzi yekupisa yekuvhunduka, iyo inogona kushatisa dema.
● Mhedzisiro paDNA uye marara marara
Kutonhora kumhepo tembiricha kunobvumira iyo cellular marara kuti isarudze, zvichiita kuti zvive nyore kupatsanura iyo DNA mumatanho anotevera. Izvi zvakare zvinobatsira kudzikamisa zviitiko zve enzyme uye zvinofambisa kubviswa kweRNA neRNARE kurapwa.
Kuwedzera kweRnase Solution
● • Chinangwa che rnase mune maitiro
Rnase Solution inowedzerwa kune degrade rna, iyo yaisakwanisa kuva co - chinatsa neDNA uye kupindira nekunyorera kwekunyora. Rnase akasarudza kugaya rna, ichisiya dna.
● Kudzivirira rna kusvibiswa
Kudzivirira rna kusvibiswa kwakakosha kune zvinoshandiswa zvinodiwa Iyo rnise kurapwa kunovimbisa kuti iyo isiriyo DNA yakasununguka kubva kuRNA zvinangwa.
Kucheneswa kweDNA
● Nzira dzekuparadzanisa DNA kubva kune mamwe ma cellular zvikamu
Nzira dzinoverengeka dzinogona kushandiswa kuchenesa DNA kubva kuLysate. Izvi zvinosanganisira phenol - chloroform chinorehwa, ethanol chairo, uye kushandisa yekutengesa e. Coli DNA kits. Imwe neimwe nzira ine zvakashata uye zvakashata, nezvikwereti zvekutengeserana zvichireva nyore uye zvinoitika.
● Kufungisisa kweDNA kuchena
Kuchena chinhu chinoshoreka chebudiriro yekunyepedzera mafomu ekuratidzira. Kitence kits, yakadai seSeller Therapy E. Colili DNA, yakagadzirirwa kubereka kumusoro - Kuchiva DNA nekubiridzira nekubvisa mapuroteni, Lipids, nezvimwe zvinosvibisa.
Kuchengetedza uye kubata kweSolated DNA
● Zvakanakisa maitiro ekuchengetedza DNA
Akambonatswa, DNA inofanira kuchengetwa mune yakakodzera buffer, kunge te buffer, at - 20 ° C kana - 80 ° C kwenguva yakareba - Nguva yekuchengetedza. Ngwarira nguva zhinji freeze - thaw come pavanogona kukonzera kusvibiswa kweDNA.
● Kuchengetedza DNA Kuvimbika Kwekudzika Kwekudzora
Kuchengetedza kutendeka kweDCA, shandisa zvisina kufanira, nucasease - ma tubes yemahara uye mhinduro. Izvi zvinovimbisa kuti DNA vanoramba vasina kusvibiswa uye zvakakodzera kuti zvikumbiro zvakadai sekuvhara, kutevedzana, uye pcr.
NezveBluekit
Jiangsu Hillgege akasimbisa dzimba dzaro (10,000㎡ gmp zvirimwa uye r & d pakati) muSuzhou, nzvimbo mbiri dzekugadzira muSheenzhen neShanghai network yekugadzira nyika. Iyo North Carolina Site muUS parizvino iri kuvakwa, nekuenderera mberi nekuparadzira kuvapo kwakawanda. Hillgee akavaka nzira yekunyora yekukudziridza cellular kurapa zvigadzirwa, kubva pakuwana kuendesa, nemapuratifomu emapuratics acid yekugadzira, Serums yekushambadzira tsika, yakavharwa yekuvandudza kukura, uye kuongororwa QC. Zvigadzirwa zvebhuruu zvakagadzirirwa kudzora kwemhando, kuona kubudirira kwesero rekurapa.
Kutumira Nguva: 2024 - 09 - 05 14 14:47:03
