Team:TUDelft/14 August 2009

(Difference between revisions)

Revision as of 16:15, 14 August 2009

First did ligation of !E and !F with the parts that were not excised from gel.

Then I did electroporation of the !E and !F gel extracted assemblies.

After that I did chemical transformation of !E and !F assemblies (both gel extracted and direct PCR restricted fragments)

oriTR_KO +L-ara +PCR and trbK_KO +L-ara +PCR backup plates made on 0.5x KAN.

Did miniprep on CB and CC culture tubes.

Plate 2, 4, 6, and 8 are overgrown. Cells survived electroporation. No colonies on knockout plates or control plates.

Ran 1.5% gel with CC and CB.

Well	Part	Expected Plasmid Size	Status
1	[http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder]	2953
2	CB	1162	? too light
3	CC	311	? SpeI not cutting

Digested CB with X + P (11uL DNA). Digested CC with E + S (4.5uL DNA).

@@ Line 65: / Line 65: @@
 Ran 1.5% gel with CC and CB.
+[[Image:Gel140809.png|600px]]
 {| border="1" align="center"
@@ Line 71: / Line 73: @@
 | 1 || [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder] || 2953 ||
 |- align="center"
-| 2 || CB || 1162 || <font color=limegreen>&#10004;</font>
+| 2 || CB || 1162 || ? too light
 |- align="center"
-| 3 || CC || 311 || <font color=limegreen>&#10004;</font>
+| 3 || CC || 311 || ? SpeI not cutting
 |}