Dzidza Sei Kuita Tae Buffer mune Zvishoma Zvishoma

Izvi Zvichishandiswa Kudzidza DNA Movement

TAE buffer ndiyo mhinduro yakagadzirwa neTris base, acetic acid uye EDTA (Tris-acetate-EDTA). Iyo inhoroondo yakawanda inonyanya kushandiswa yakashandiswa agarose gel electrophoresis mune kuongorora kweDNA michina inokonzerwa nekugadzirisa kwePCR , zvirongwa zvekuchenesa DNA kana kuti DNA kuongorora maitiro.

Ichi chine simba ine simba ineononi uye yakaderera kugadzirisa simba. Izvo zvakanakisisa zvakakosha ku electrophoresis yepamusoro (> 20 kb) zvidimbu zveDNA uye inoda kuchinjwa kazhinji kana kuidzoswa kwenguva yakareba (> 4 h) gel rinowedzera nguva.

Uine izvozvo mupfungwa, iwe ungada kuongorora mabheji akawanda emutambo.

Tichifunga kuti chigadziro chiri nyore kuita uye matanho anogona kuitwa nokukurumidza, kuita batch imwe chete panguva imwe hazvifaniri kunyanya kupedza nguva kana kuti zvakaoma kubvisa. Uchishandisa mirayiridzo iri pasi apa, inofanira kutora maminitsi makumi matatu chete kuti uite TAE. Heino nzira:

Gadzirira Solution Solution ye EDTA

An EDTA (ethylenediamine tetraacetic acid) sarudzo inogadzirirwa mberi kwenguva. EDTA haizombopindi mhinduro zvachose 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). Dissolve mu 400-milliliters mvura yakasanganiswa uye gadzirisa pH ne sodium hydroxide (NaOH). Tambanudza kumusoro kwemugadziro wekupedzisira kwema 500 milliliters.

Kusimudza A Stock Solution ye TAE

Ita yakanyatsotarisa (50x) yekugadzirisa shanduko ye TAE nekuyera masendimita 242 eTris base (FW = 121.14) nekuipedza iyo inenge 750 mamiriyoni emvura yakasiyiwa.

Ngwarira kuwedzera 57.1 milliliters ye glacial acid uye 100 mamiriyoni e 0.5 M EDTA (pH 8.0). Mushure meizvi, shandurai mhinduro kumugumo wokupedzisira we 1 litre. Izvi zvinogadziriswa zvinogona kuchengetwa pamusana wekupisa. Iko pH yeiyo mutsara haina kuchinjwa uye inofanira kuva ne8,5.

Gadzirira Kuita Solution ye TAE

Shanduro yekushanda ya 1x TAE buffer inogadzirwa nokungoshandura zvigadziriswa zvayo nema 50x mumvura yakasvibiswa.

Mafungiro ekupedzisira mashoma ndeomamiriyoni 40. Tris-acetate uye 1 mM EDTA. Iyo mhuka yakagadzirira ikozvino kushandiswa mukugadzira gel agarose.

Izvo Zvaunoda Kuti Uve Mutsva

Kubvira pakugadzira mutengo we TAE kunongoda kuiswa kwekukurumidza uye nyore nyore mirayiridzo, nhamba yezvinhu zvaidiwa nokuda kwayo haisi kunyanya. Iwe unongoda EDTA munyu wemidium, Tris base, uye glacial acetic acid.

Kuita chibvumirano chinodawo pH mamita uye maitiro ekugadzirisa, sezvakakodzera. Iwewo unoda 600 milliliter uye 1500 milliliters beaker kana mabhasiki pamwe nekupedza zvidzidzo. Pakupedzisira, iwe unoda mvura yakasvibiswa, bhutiro, uye inoputira mahwendefa.

Wrapping Up

Iwe unoronga kupi kugadzira chikwata - panzvimbo yechikoro, nzvimbo yebasa, kana imwe nzvimbo, yakafanana neimba yako? Chengetedza zvinyorwa usati watanga kuve nechokwadi kuti une zvinhu zvose zvakasarudzwa zveTAT buffer.

Kana iwe uchida haufaniri kubhadhara zvinhu, kusarudza kuita chigadziri kunze kwemusha wako kunokurudzirwa. Iwe unogona kurongedza nyore nyore zvinhu zvaunoda paIndaneti kana kushanyira imwe nzvimbo yezvigadzirwa.