The printable version is no longer supported and may have rendering errors. Please update your browser bookmarks and please use the default browser print function instead.
Karmella's BioBrick Cloning
|
<html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>
|
01/10/13
- Violacein: successful growth; re-streak
- Golden gate/ transformation: transform DH5α-Turbo with gg assemblies from 1/09/13
- Golden gate trial 2: new PCR primers
Violacein
- Plate 1: Kan plasmid
- Plate 2: Amp plasmid (violacein exp.)
Golden gate trial 1
- Add total 10.0 μL run to 50 μL DH5α-Turbo; incubate on ice 10 min.; plate on amp agar
Golden gate trial 2
- Re-designed primers to include "handle" preceding the BsmBI sites and a spacer between BsmBI and a 4 bp overlap region
Reagent
|
Volume
|
Expected: 1. pSB1A3 = ~2000 2. hPCD = 186 3. BL01 = 2520
|
|
DNA(plasmid) |
0.2 μL
|
primer 1 |
1.0
|
primer 2 |
1.0
|
2x GoTaq |
25.0
|
dH2O |
22.8
|
|
50.0 μL
|
PCR
- 95°C, 3 min.
- [95°C, 3 min.; 95°C, 3 min.; 95°C, 3 min.] x35
- 72°C, 3 min.
- 4°C, ∞
Sample
|
OD 260
|
260/280
|
ng/μL
|
1. pSB1A3 |
0.126 |
1.888 |
77.357
|
2. hPCD |
0.069 |
2.192 |
17.025
|
3. BL01 |
0.121 |
1.898 |
66.878
|
|