Baylee Murphy- 11/20/15 Session 3
KAH126- MV2 Culture inoculation results from Session 2
- Liquid cultures 1 and 2 both had cloudy yellow broth, indicating growth
- Streaks for both showed colony growth
Samples for mini prep and analysis
- (1) Sample 1- KAH126-MV2 colony 1, [BB#}; part = [length in bp]; vector= [length in bp]
- (2) Sample 2- KAH126-MV2 colony 2, [BB#}; part = [length in bp]; vector= [length in bp]
DNA concentration Data
| Sample Read#
|
Location
|
260 Raw
|
280 Raw
|
320 Raw
|
260
|
280
|
260/280
|
ng/µL
|
| BM KAH126-1 |
C2 |
0.32 |
0.207 |
0.08 |
0.249 |
0.131 |
1.909 |
249.344
|
| BM KAH126-1 |
C3 |
0.313 |
0.201 |
0.074 |
0.245 |
0.128 |
1.911 |
244.882
|
| BM KAH126-2 |
D2 |
0.323 |
0.203 |
0.06 |
0.272 |
0.146 |
1.861 |
272.416
|
| BM KAH126-2 |
D3 |
0.319 |
0.2 |
0.066 |
0.258 |
0.136 |
1.904 |
258.244
|
Plasmid Validation
Restriction Digest Table
- Checked plasmid minipreps with Xba digest
| Reagent
|
Volume
|
Expected:
1. KAH126-MV2 = 1934 bp and 4505 bp
|
10 μL/lane; 1% agarose; Ladder
|
| DNA(plasmid) |
1.0 μL
|
| 10X buffer |
1.5 μL
|
| Xba |
1.0 μL
|
| dH2O |
10.5 μL
|
| |
10 μL --> 37°C/ 20 min.
|
Control for 1 and 2
| Reagent
|
Volume
|
| DNA(plasmid) |
1.0 μL
|
| 10X buffer |
1.5 μL
|
| dH2O |
11.5 μL
|
Results of restriction digest show expected bands and indicate Xba enzyme worked
KAH126 incluswa Hpdc, scar, mcherry: about 1000 bp
- forward primer- DD122 10 uM
- reverse primer-DD123 10 uM
DNA Sequencing Samples
- Submitted to DNASU on: '11/20/15'
- Order Number:
- Sample 1- KAH126-MV2 colony 1, forward primer DD122
- Sample 2- KAH126-MV2 colony 1, reverse primer DD123
- Sample 3- KAH126-MV2 colony 2, forward primer DD122
- Sample 4- KAH126-MV2 colony 2, reverse primer DD123
|
Haynes Lab Cloning Bootcamp
