Paul.xu at 14:15, 25 June 2017

2017-06-25T14:15:04Z

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Composition of 1x TAE buffer  ■ 40 mM Tris (pH 7.6)  ■ 20 mM acetic acid  ■ 1 mM EDTA 

Preparation of 50x TEA stock solution 

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

 Composition of 1x TBE buffer

■ 89 mM Tris (pH 7.6) ■ 89 mM boric acid ■ 2 mM EDTA

Preparation of 10x TBE stock solution

To prepare 1 liter of 10× TBE dissolve following components in 600 ml deionized water: ■ 108 g Tris base (FW = 121) ■ 55 g boric acid (FW = 61.8) ■ 40 ml 0.5 M EDTA (pH 8.0) 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 : [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4021863/ https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4021863/]
[[Category:Buffer Recipe]]

Buffer for Agarose Gel Electrophoresis - Revision history

Paul.xu at 14:15, 25 June 2017