DNA Electrophoresis Common Issues

Gel electrophoresis ndi imodzi mwa njira zazikulu zomwe zimagwiritsidwa ntchito mu biology ya molekyulu posanthula DNA.Njira imeneyi imaphatikizapo kusamuka kwa tizidutswa ta DNA kudzera mu gel, kumene timapatukana potengera kukula kapena mawonekedwe.Komabe, kodi munayamba mwakumanapo ndi zolakwika zilizonse pakuyesa kwanu kwa electrophoresis, monga zopaka zopaka pa gel osakaniza, kapena palibe mabandi pa gel osakaniza?Kodi cholakwikacho chingakhale chiyani?

zokwiyitsa

Akatswili athu afotokozera mwachidule mabanja angapo azovuta apa kuti muwafotokozere.

1. Zopaka zopaka pa gel osakaniza agarose

kupaka bandeji pa gel osakaniza

DNA yawonongeka.Pewani kuipitsidwa ndi ma nuclease.

● Chombo cha electrophoresis sichatsopano.Pambuyo pakugwiritsa ntchito mobwerezabwereza electrophoresis buffer, mphamvu ya ionic imachepa, ndipo pH mtengo wake umawonjezeka, kotero mphamvu ya buffer imafooka, zomwe zimakhudza mphamvu ya electrophoresis.Ndikofunikira kuti m'malo mwa electrophoresis buffer pafupipafupi.

● Mikhalidwe yolakwika ya electrophoresis inagwiritsidwa ntchito.Musalole kuti magetsi apitirire 20 V / cm, ndipo sungani kutentha kwa <30 ° C pa electrophoresis.Kwa chimphona chachikulu cha DNA strand electrophoresis, kutentha kuyenera kukhala kosachepera 15 ° C. Yang'anani kuti electrophoresis buffer ili ndi mphamvu zokwanira.

● DNA yochuluka kwambiri inayikidwa pa gel.Chepetsani kuchuluka kwa DNA.

● Mu DNA muli mchere wambiri.Gwiritsani ntchito mpweya wa ethanol kuti muchotse mchere wochuluka kwambiri.

● DNA inali ndi mapuloteni.Gwiritsani ntchito zowonjezera za phenol kuchotsa mapuloteni apamwamba.

● DNA inasinthidwa.Musati kutentha pamaso electrophoresis.Sungunulani DNA mu buffer ndi 20 mM NaCl.

2. Anomalies DNA band kusamuka

● Kukonzanso kwa tsamba la COS lachidutswa cha λHind III.Tenthetsani DNA kwa mphindi 5 pansi pa 65 ° C pamaso pa electrophoresis, kenaka muziziritse pa ayezi kwa mphindi zisanu.

● Mikhalidwe yolakwika ya electrophoresis inagwiritsidwa ntchito.Musalole kuti magetsi apitirire 20 V / cm, ndipo sungani kutentha kwa <30 ° C pa electrophoresis.Onani buffer ya electrophoresis ili ndi mphamvu zokwanira zotchingira.

● DNA inasinthidwa.Musati kutentha pamaso electrophoresis.Sungunulani DNA mu buffer ndi 20 mM NaCl.

3. Mabandi a DNA okomoka kapena osapezeka pa gel osakaniza

ma DNA ofooka

● Panalibe unyinji wokwanira kapena kuchuluka kwa DNA yopakidwa pa gel.Wonjezerani kuchuluka kwa DNA.Polyacrylamide gel electrophoresis ndizovuta kwambiri kuposa agarose electrophoresis, ndipo kutsitsa kwachitsanzo kumatha kuchepetsedwa moyenera.

● DNA inawonongeka.Pewani kuipitsidwa ndi ma nuclease.

● DNA inajambulidwa ndi electrophoresi kuchokera mu gel.Electrophorese gel kwa nthawi yochepa, gwiritsani ntchito magetsi otsika, kapena gwiritsani ntchito gel okwera kwambiri.

● Gwero la kuwala kwa W lolakwika linagwiritsidwa ntchito powonera DNA ya ethidium bromide-stained DNA.Gwiritsani ntchito kuwala kwafupipafupi (254 nm) W kuti mumve zambiri.

4. Magulu a DNA akusowa

DNA yaying'ono yaying'ono idapangidwa ndi electrophoresis kuchokera pa gel.Electrophorese gel kwa nthawi yochepa, gwiritsani ntchito magetsi otsika, kapena gwiritsani ntchito gel okwera kwambiri.

● N'zovuta kusiyanitsa magulu a DNA a ma molekyulu ofanana.Onjezani nthawi ya electrophoresis, ndikuyang'ana ndendegel osakaniza kuti atsimikizire kuti gel osakaniza agwiritsidwa ntchito moyenera.

● DNA inasinthidwa.Musati kutentha pamaso electrophoresis.Sungunulani DNA mu buffer ndi 20 mM NaCl.

● Zingwe za DNA ndi zazikulu, ndipo ma electrophoresis a gel ochiritsira si oyenera.Unikani pa pulse gel electrophoresis.Ndi zinthu zina ziti zomwe mwakhala nazo ndi agarose gel electrophoresis?Tidzafufuza zambiri za maupangiri mtsogolomo.

Beijing Liuyi Biotechnology Co., Ltd (Liuyi Biotech) ndi kampani yapadera yomwe imayang'ana kwambiri zinthu zokhudzana ndi electrophoresis ku China.Nkhani yake imayamba mu 1970 pomwe China inali isanalowe mukusintha ndikutsegula nthawi.Kupyolera mu chitukuko cha zaka, Liuyi Bitotech ili ndi mtundu wake, womwe umadziwika kuti Liuyi Brand pamsika wapakhomo wa mankhwala a electrophoresis.

Mtundu wa Liuyi uli ndi mbiri yopitilira zaka 50 ku China ndipo kampaniyo imatha kupereka zinthu zokhazikika komanso zapamwamba padziko lonse lapansi.Kupyolera mu chitukuko cha zaka, ndizoyenera kusankha kwanu!

Maselo opingasa a electrophoresis (matangi/zipinda) a Liuyi Biotech ndi apamwamba kwambiri komanso mawonekedwe abwino.Ndi makulidwe osiyanasiyana a ma tray a gel, amatha kukwaniritsa zomwe mukufuna.Tili ndi gulu lathu luso ndi fakitale.Kuchokera pakupanga mpaka kupanga, zopangira mpaka magawo ofunikira, titha kuwongolera njira yonse.Mndandanda wa DYCP 31 ndi wa DNA electrophoresis, omwe ndi chitsanzoChithunzi cha DYCP-31BN, Chithunzi cha DYCP-31CN,Chithunzi cha DYCP-31DN,ndiDYCP-31E.Kusiyanitsa pakati pawo ndi kukula kwa gel ndi mtengo.Timapereka kukula kwazinthu zonse kwa makasitomala athu.ChitsanzoDYCP-32Cakhoza kupanga gel osakaniza 250mm * 250mm.

1-1

Pakalipano, tikupangira mphamvu zathu za electrophoresisDYY-6C,DYY-6DndiDYY-10Ckwa maselo athu a electrophoresis (akasinja / zipinda) DYCP-31 ndi 32 mndandanda.

1-4

Ngati mukufuna zambiri za malonda, chonde pitani patsamba lino kuti mumve zambiri, ndikulandilidwa kuti mutitumizire imelo kuti tidziwe zomwe mukufuna, ndikuwona ngati titha kukupatsirani mayankho.

Kuti mudziwe zambiri za ife, chonde titumizireni imelo[imelo yotetezedwa], [imelo yotetezedwa].


Nthawi yotumiza: May-09-2022