Monday, June 12, 2006

Restriction Digestion

Gel shown are cultures 1, 2, 3 and 31 on a 1.3% TAE gel stained with Sybr Safe. Flanking lanes are Bio-Rad's 500 bp ladder (16 fragments from 500 to 8000 every 500 bp). Gel 2 is cultures 32, 33, 34 and 35, not shown.

I'm now working at home since school has ended. Over the past week I've done minipreps on eight cultures (grown in Teriffic Broth) and digested with Bgl II. Miniprep system is Sigma's GenElute Plasmid kit.

Calculated fragments
Correct at 557 and 4335
Reverse Orientation at 804 and 4088
No Insert at 3653

Clone #31 (fifth column shown at right) has about the correct size fragments, at approximately 4280 and 534. As far as I know, that is close enough to assume correct. My error probably comes from the fact that the bands were fairly wide on the gel image. Clones 2 and 3 have faint bands at 4280, and given that the heavy band is brighter, it might be safe to assume that the 557 bp band is present but too faint to see.

Two cultures had one band at about 2500 bp. I'm not sure what those bands were.


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