Team:Groningen/Labwork/22 July 2013

From 2013.igem.org

(Difference between revisions)

Revision as of 12:46, 22 July 2013

Mirjam
Made a PCR for silk 2 and 3 (strepF-R and F-strepR)in triplo
- 28 ul MQ
- 1.5 ul DMSO
- 10 ul GC buffer
- 1 ul of each dNTP
- 2.5 ul primer F
- 2.5 ul primer R
- 0.5 ul phusion
- 1 ul genomic DNA (BBa_...016)

Run a gel with the products to determine if the silk is present. Silk is present in high amounts, but only for the parts under 500 bp. That is why a new PCR is made for both silks.

Run a gel with the ligation product of silk and the PCR afterwards (made on 19-07). Figured out why the ligation did not work.

Made a new restriction digest for silk without strep tag.

Inne
Ran Gel to check everything that was in the freezer before discarding it.
Gel was 1.5% and ran at 90V for 45 min

Checked tubes:

Slot	Description Lid	Description Side
1	1kb generuler	Fermentas 1kb DNA Ladder (and more)
2	1	Silk 4.1ng/uL Strep R-F
3	1	Silk 4.1ng/uL F-R
4	2	Silk 4.6ng/uL Strep R-F
5	2	Silk 4.3ng/uL Strep R-F
6	3	Silk 4.9ng/uL R-strepF
7	3	Silk 5.9 ng/uL R-strepF
8	Silk without strep tag	10-07-13 Silk without strep tag 5.1 ng/uL
9	2	16-07 Silk
10	3	16-07 Silk

Sander
did a ligation reaction on silk 1 with two of the Signal Sequences (MotB and EstA) at 1:1 ratio.
ran a gel with 0.8% agarose at 90V for 34 min.
no bands visible but external factors migh be influencinfg imaging zo samples were stored in freezer (box TEMP: marked on top with M-S and E-S on side with lig m-silk and lig e-silk)

@@ Line 78: / Line 78: @@
 </tr>
 </table>
+'''Sander'''
+<br> did a ligation reaction on silk 1 with two of the Signal Sequences (MotB and EstA) at 1:1 ratio.
+<br> ran a gel with 0.8%  agarose at 90V for 34 min.
+<br>no bands visible but external factors migh be influencinfg imaging zo samples were stored in freezer (box TEMP: marked on top with M-S and E-S on side with lig m-silk and lig e-silk)