Lab Aug 25 2009

Miniprep: Qiagen Miniprep Protocol

2 K235000 one from each plate

1 K235001

2 K235002 one from each plate

Insertion digest of K145303 to put into Chl backbone, and upstream digest of same to combine with ribokey part.

Put the competent cells in room temp shaker at 9.30 am with OD of 0.05

Agarose Gel Electrophoresis Protocol

Gel 1 started at 12.22 pm Ladder K235000 100 uL K235000 200 uL K235001 100 uL K235002 100 uL K235002 200 uL K145303 K081005

Band

present Band Inconsistent Band *Multiple Bands Band Inconsistent Band Band No band Size 1441 2213

11057?

7833

5635

3783

2871 1913 1745 2130

Aug 25 gel 1.tif

-Layne

This lane showed various faint bands, a normal strength band at 7833 and a super intense band at 3783. Has it been cotransformed with multiple plasmids as Chris suggested?

You can look at my calculations here.

Assemblies: 3A Assembly Protocol

J23102+P1010Kan = K235006

K145303+P1010Kan = K235007

K145303+K235001+P1010Cm = K235008

Digested, ligated and transformed: Transformation Protocol

Gel 2 Ladder Size 2308 No band

4045

2959 No band 2776 No band No band

aug 25 gel 02.tif

-Layne

Hopefully someone knows what was in this gel?

This was a really blurry gel, we can't put much weight on the values from it.

You can look at my calculations here.