Reagents used...in restriction digestion and electrophoresis - (Mar/14/2006 )

Hi,

I was wondering if anyone can give me a hand here...

1) Why is restriction buffer always added before adding restriction enzymes? (I thought it was to
provide a medium for the enzymes, but don't know if it is true)

2) What's the function of TAE buffer?

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1. Buffer is often in 10x concentration, its better to dilute it to apprx. 1x concentration with water then at last adding enzyme. Re's don't like buffers in to high concentration or "pure" water.
2. Tae is for electrophoresis, buffering for suitable pH and having the ions for the circuit.

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moreover, bad pH or bad salt concentration may drive non specific cutting of RE.

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