Izvi zvinoshandiswa kwe electrophoresis kuparadzana kweDNA
Chirwere cheTBE chinonyanya kukosha pakuparadzanisa zviduku zviduku zveDNA (MW <1000), zvakadai semidziyo miduku yekudzivirira inyme enzyme digest.
TBE ine dambudziko guru rekutengesa uye ichapa sarudzo yakasimba kudarika taE TAE . TAE buffer ndiyo mhinduro yakagadzirwa neTris base, acetic acid uye EDTA (Tris-acetate-EDTA).
TBE inowedzera inodhura kupfuura TAE uye inovhara DNA ligase, iyo inogona kukonzera matambudziko kana kutevera DNA kucheneswa uye kugadzirisa matanho kunangwa. Nezviito zvitatu zvinotevera zvinotevera, dzidza kuti ungaita sei TBE. Izvo hazvifaniri kutora zvinopfuura maminitsi makumi matatu kuti zvigadzire dambudziko iri. Heino nzira:
Gadzirira Solution Solution ye EDTA
EDTA (ethylenediamine tetraacetic acid) sarudzo inofanira kugadzirirwa mberi kwenguva. EDTA haizoperi zvachose muganhu kusvikira pH ichinjwa kusvika 8.0. Kwema 500-milliliter stock solution ye 0.5 M EDTA, inofanidza 93.05 gramu ye EDTA saldium disodium (FW = 372.2). Zvadaro gurutsa mu 400 milliliters mvura yakasanganiswa uye gadzirisa pH neNaOH. Mushure mokudaro, sarudza kumusoro kwegadziriro yemugumo wokupedzisira wemamiriyoni 500.
Gadzirira Solution Solution yeTBE
Ita yakagadziriswa (5x) mugadziriso weTBE nekuyera 54 g g Base reTris (FW = 121.14) uye 27.5 g boric acid (FW = 61.83) uye kuputsa zvose mumakriyoni anenge 900 emvura yakakanganiswa. Zvadaro wobva wawedzera 20 millilitres e 0.5 M EDTA (pH 8.0) uye gadzirisa chinogadziriswa kumugumo wokupedzisira we 1 litre.
Iyi sarudzo inogona kuchengetwa pamusana wekupisa kwemahombekombe asi kunonoka kuchagadzirisa muzvigadzirwa zvekare. Chengetedza mutsara mumabhodhoro egirazi uye uregere kana pakaitika.
Gadzirira Kushanda Solution yeTBE
Nokuda kwe agarose gel electrophoresis, chigadziro cheTBE chinogona kushandiswa pamusana we 0.5x (1:10 dilution yeganda rakaiswa). Deredza mukana wekugadzirisa zvinokonzerwa ne10x mumvura yakasvibiswa. Mafungiro ekupedzisira akawandisa ndiwo mamita 45MM Tris-borate uye 1 mM EDTA. Iyo mhuka yakagadzirira ikozvino kushandiswa mukugadzira gel agarose .
Zvaunoda
Kuti uite TBE tape, iwe unoda zvinhu zvina chete. Zvinhu zvakasara pane urongwa izvi zviyero. Izvo zvinhu zvina zvinodiwa ndeye EDTA munyu wemidium, tsvina yeTris, boric acid uye mvura yakakosha.
Kana iri midziyo, iwe uchada pm mamita uye maitiro ekugadzirisa, sezvakakodzera. Mukuwedzera kune izvozvo, iwe unoda 600-milliliter uye 1500-milliliter beaker kana flasks. Kurongedza zvipfeko zvako zvinoshandiswa zvinopedza zvidzidzo uye mapurisa ekuputira uye mapuraneti anotema.
Tarisa kuongorora pabazi raunenge uri kushandisa kuti uve nechokwadi chokuti une zvose zvaunoda usati watanga. Hapana chakaipa kudarika kumira pakati pekugadzirisa mhinduro nokuti wakabuda kunze kwezvinhu zvakakodzera.
Kana basa rako riri kuchikoro kana nzvimbo yako yebasa, tarisa nevashandi vakakodzera kuti vaone kuti vane zvinhu zvose zviri muchikwata. Kuita izvi kunogona kukuponesa nguva nemasimba pakupedzisira.