Day 3- Minipreps
Liquid Culture Results
NOTE: Cultures and Streaks taken out after 11 hours (37 incubator) and left on lab bench for 11 hours.
hPCD
- Cultures Innoculated 09/12/13
- Cultures 1 and 2: Both show growth
- Streak 1 and 2: Both show growth
Ubc
- Cultures Innoculated 09/12/13
- Cultures 1 and 2: Both show growth
- Streak 1 and 2: Both show growth
Samples for Minipreps and Analysis
- Sample 1= hPCD/V0120 colony 1; BBa_J176000; part= 186 bp; vector= 4968 bp
- Sample 2= hPCD/V0120 colony 2; same
- Sample 3= Ubc/V0120 colony 1; BBa_J176003; part= 368 bp; vector= 4968 bp
- Sample 4= Ubc/V0120 colony 2; same
- Sample 5= hPCD/V0120; BBa_J176000; part= 186 bp; vector= 4968 bp
- Sample 6= Ubc/V0120; BBa_J176003; part= 368 bp; vector= 4968 bp
DNA Concentration Data
| Plasmid |
OD260 |
OD260/280 |
ng/μL
|
| 1. hPCD 1 |
0.279 |
1.831 |
278.676
|
| 2. hPCD 2 |
0.242 |
1.834 |
241.754
|
| 3. Ubc 1 |
0.097 |
1.787 |
96.614
|
| 4. Ubc 2 |
0.146 |
1.789 |
145.548
|
Restriction Digest Table
- Checked plasmid minipreps with EcoRI/PstI digests
| Reagent
|
Volume
|
Expected:
1. hPCD = 186 bp, 4968 bp
2. Ubc = 368 bp, 4968 bp
|
15 μL/lane; 1% agarose; Ladder
|
| DNA(plasmid) |
3.0 μL
|
| 10X FD Buffer |
1.5
|
| EcoRI |
1.0
|
| PstI |
1.0
|
| dH2O |
8.5
|
| |
15 μL --> 37°C/ 15 min.
|
Conclusion
There were definite gel lines at ~4900 and faint lines at ~200 for hPCD (lanes 1,2,5) and ~ 350 for Ubc (3,4,6).
This signifies a working gel.
DNA Sequencing Samples
- Submitted to DNASU on: '8/14/13'
- Order Number: 8043
- Plasmid 1 (hPCD) - Forward P001
- Plasmid 1 (hPCD) - Reverse P002
- Plasmid 4 (Ubc) - Forward P001
- Plasmid 4 (Ubc) - Reverse P002
|
Haynes BioBrick Cloning
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