Le ntlanzi isetyenziselwa ukuFunda i-DNA Movement
Esi sikhonkco sinamandla amakhulu e-ionic kunye ne-low buffering capacity. Kuyafaneleka ukuba i-electrophoresis yamacandelo amakhulu (> 20 kb) e-DNA kwaye iya kufuneka ukuba ithatyathelwe ngokuphindaphindiweyo okanye iphindwe ixesha elide (> 4 h).
Ngaloo nto engqondweni, unokuba ufuna ukucubungula amanqwanqwa amathandathu.
Ngenxa yokuba i-tamper ilula ukwenza kwaye amanyathelo angenziwa ngokukhawuleza, ukwenza i-batch engaphezu kweyodwa ngexesha akufanelekanga ngokukhawuleza ixesha okanye kunzima ukukhupha. Ukusebenzisa imiyalelo engezantsi, kufuneka kuthathe imizuzu engama-30 ukwenza i-TAE buffer. Nantsi indlela:
Lungisa iSolution Solution ye-EDTA
Isisombululo se-EDTA (i-ethylenediamine tetraacetic acid) isilungiselelwe ngaphambi kwexesha. I-EDTA ayiyi kufikelela kwisisombululo kuze kube yilapho i-pH ishintshwe malunga ne-8.0. Isisombululo se-500-milliliter yesitoreji se-0.5 M EDTA, isilinganise i-93.05 amagremu we-EDTA ityuwa ye-disodium (FW = 372.2). Dissolve ku-400-milliliters amanzi adibeneyo kwaye utshintshe i-pH nge-sodium hydroxide (i-NaOH). Phakamisa isisombululo somlinganiselo wokugqibela wama-milliliters ama-500.
Ukukhupha isisombululo seStock ye-TAE
Yenza isisombululo esisisigxina (50x) se-TAE ngokulinganisa i-242 grams ye-Tris base (FW = 121.14) kunye nokuyichitha malunga nama-milliliters angama-millilitha amanzi.
Yongeza ngokucophelela i-57.1 milliliters ye-glacial acid kunye ne-100 milliliters ye-0.5 M EDTA (pH 8.0). Emva koko, hlengisa isisombululo kwisixa sokugqibela se 1 litre. Isisombululo sesisitya singagcinwa kwiqondo lokushisa. I-pH yale khowudi ayilungiswanga kwaye kufuneka ibe malunga ne-8.5.
Lungiselela isisombululo sokuSebenza se-TAE
Isisombululo sokusebenza se-tx 1x TAE senziwa ngokululaza isisombululo se-stock nge-50x emanzini afakwe ngamanzi.
Iimigangatho zokugqibela ze-solu zi-40 mM. I-Tris-acetate kunye ne-EDM eyi-1 yemitha. I-buffer manje ilungele ukusetshenziswa kwi-gel agarose.
Okukufunayo kwiBampu
Ekubeni ukwenza i-TAE buffer idinga kuphela imiyalelo elula kwaye elula, inani lezinto ezifunekayo kulo alikho. Uza kufuna nje ityuwa ye-disodium, i-Tris base, kunye ne-glacial acetic acid.
Ukwenza i-buffer nayo idinga imitha ye-pH kunye nemigangatho yokulinganisa, ngokufanelekileyo. Uya kufuneka kwakhona ama-600 milliliter kunye ne-1500 milliliters beakers okanye iiflasks kunye nezilwanyana eziphunyeziweyo. Ekugqibeleni, uzakufuna amanzi adibeneyo, ugxobhoze imivalo, kwaye ugubungele iiplate.
Ukuqhawula phezulu
Uceba phi ukwenza i-buffer-kwindawo yesikolo, indawo yomsebenzi, okanye kwenye indawo, njengekhaya lakho? Hlola i-inventory phambi kokuba uqale ukuqinisekisa ukuba unazo zonke izixhobo ezingentla apha ze-TAE.
Ukuba ngaba unganakho ukuhlawula izixhobo, ukhethe ukwenza i-tampon ngaphandle kwekhaya lakho kucetyiswa. Unokwenza lula ukuba ulungiselele izinto ozifunayo kwi-intanethi okanye u tyelele ivenkile ekhethekileyo.