Week of 5/31

From 2010.igem.org

(Difference between revisions)

Revision as of 18:00, 29 July 2010

Flasks Mini prepped:

   - B0010 (Transcriptional Terminator)
   - R0062 (Promoter activated by LuxR in concert with HSL)
   - C0012 (lacI repressor from E. coli)
   - R0010 (promoter (lacI regulated))
   - C0051 (cI repressor from E. coli phage lambda)
   - B0034 (RBS)

All of the mini preps were digested with the following enzymes:

   - B0010 (EX)
   - R0062 (EX)
   - C0012 (XP)
   - R0010 (XP)
   - C0051 (XP)
   - B0034 (XP)
   - PCI   (XP)

E=EcoRI , X=XBaI , P=PstI , S=SpeI

To finish the digest, it was placed in a water bath at 37 degrees Celsius for 1 hour. The next step was to heat kill the reaction, so we placed the digest products into an 80 degrees Celsius water bath for 20 minutes.

Following the protocol we ran the digested products on an Agarose gel for ~45 minutes.

As we can see PCI did not show up but C0012 and C0051 showed up and the bands were extracted and then gel purified. B0034 and R0010 did not however show up on the gel. We were expecting bands further down with base pairs of around 500. The gel purifications were then ligated and then later transformed. After transforming and waiting for an hour, the transformations were plated and put in the 37 degrees incubator.

@@ Line 20: / Line 20: @@
 *E=EcoRI , X=XBaI , P=PstI , S=SpeI
-* After the digest was put in the 37 degrees water bath for an hour and then heat killed in the 80 degrees water bath, the digests were then ran on a gel. The picture of the gel was taken and added below.
+* To finish the digest, it was placed in a water bath at 37 degrees Celsius for 1 hour. The next step was to heat kill the reaction, so we placed the digest products into an 80 degrees Celsius water bath for 20 minutes.
+*Following the protocol we ran the digested products on an Agarose gel for ~45 minutes.