Editing Buffer for Agarose Gel Electrophoresis

<br/>Composition of 1x TAE buffer&nbsp;<br/>■ 40 mM Tris (pH 7.6)&nbsp;<br/>■ 20 mM acetic acid&nbsp;<br/>■ 1 mM EDTA&nbsp;

Preparation of 50x TEA stock solution&nbsp;

To prepare 1 liter of 50× TAE dissolve following components in 600 ml of deionized water:&nbsp;<br/>■ 242 g Tris base (FW = 121)&nbsp;<br/>■ 57.1 ml glacial acetic aci&nbsp;<br/>■ 100 ml 0.5 M EDTA (pH 8.0)

<br/>Composition of 1x TBE buffer

■ 89 mM Tris (pH 7.6)<br/>■ 89 mM boric acid<br/>■ 2 mM EDTA

Preparation of 10x TBE stock solution

To prepare 1 liter of 10× TBE dissolve following components in<br/>600 ml deionized water:<br/>■ 108 g Tris base (FW = 121)<br/>■ 55 g boric acid (FW = 61.8)<br/>■ 40 ml 0.5 M EDTA (pH 8.0)<br/>Adjust final volume to 1 liter with deionized water.



Use 0.5 X TAE or TBE for electrophoresis

[[File:TAE TBE vs TA TB.jpg]]



[[File:TAE Vs TBE.jpg]]

[[File:TB buffer.jpg]]

Standard 1 x TBE buffer produces sharp bands and good resolution in gels run at typical low voltages of ~ 10 V/cm. However, such gels usually require run times of 50 – 90 min to achieve optimum separation of DNA fragments

TBE: good resolution

Use of 0.5 x TB also produced good separation of bands at voltages up to 250 V, though resolution was compromised at 300 V

Gels prepared and run in 0.3 x TB exhibited low resolution and compression of bands toward each other at all voltages, with strongest compression and non-uniform band intensities apparent at 250 and 300 V. Thus, good resolution was achieved using both 0.5 x TB and 1 x TB at voltages up to 250 V (25 V per cm), though all bands were closer to each other at the higher voltages. EDTA is not essential for electrophoresis and is added as a preventative to inhibit DNA nucleases

ref&nbsp;: [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4021863/ https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4021863/]
[[Category:Buffer Recipe]]