"
Team:DTU-Denmark/Notebook/27 June 2013
From 2013.igem.org
(Difference between revisions)
(→208 lab) |
(→208 lab) |
||
| Line 9: | Line 9: | ||
==Procedure== | ==Procedure== | ||
Signal peptides on 4% gels 80V for 45 min. All other PCR-products on a 1% gel 100V for 45 min. | Signal peptides on 4% gels 80V for 45 min. All other PCR-products on a 1% gel 100V for 45 min. | ||
| - | Two new PCRs with ramp from 70°C → 60°C and 60°C → 50°C. Sec signal peptide on 70°C → 60°C and TAT signal peptide with new primer set 2 and 3 respectively on program 60°C → 50°C. | + | Two new PCRs with ramp from 70°C → 60°C and 60°C → 50°C. Sec signal peptide on 70°C → 60°C and TAT signal peptide with new primer set 2 and 3 respectively on program 60°C → 50°C. Also backbone, GFPs and RFP where on these programs but non worked. |
[[File:27.06.13_Sec_and_TAT_signalP.jpg|thumb|left|The gel of the successful PCR-amp. of the two signal peptides]] | [[File:27.06.13_Sec_and_TAT_signalP.jpg|thumb|left|The gel of the successful PCR-amp. of the two signal peptides]] | ||
| + | Purification af the signal peptides where done with Illustra gfx G50 with filters all sequences under 50 bp. | ||
| + | Gel purification of weak band of the GFP SF. | ||
== Conclusion from today == | == Conclusion from today == | ||
| + | We have all PCR-fragments except the backbone. | ||
Revision as of 07:29, 28 June 2013
Contents |
208 lab
Main purposes today
Run gels to verify yesterdays PCR-prodcuts. Make new PCR to amplify the signal peptides and the backbone.
who were in the lab
Gosia, Henrike, Kristian
Procedure
Signal peptides on 4% gels 80V for 45 min. All other PCR-products on a 1% gel 100V for 45 min. Two new PCRs with ramp from 70°C → 60°C and 60°C → 50°C. Sec signal peptide on 70°C → 60°C and TAT signal peptide with new primer set 2 and 3 respectively on program 60°C → 50°C. Also backbone, GFPs and RFP where on these programs but non worked.
Purification af the signal peptides where done with Illustra gfx G50 with filters all sequences under 50 bp. Gel purification of weak band of the GFP SF.
Conclusion from today
We have all PCR-fragments except the backbone.
