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Revision as of 03:02, 7 September 2016
Week 10: 8/29 - 9/4
8/29
AmilCP BioBrick Digests
Content | Experimental ul | Control ul |
---|---|---|
H2O | 46.5 | 47.0 |
Cutsmart | 5.0 | 5.0 |
XbaI | 0.5 | X |
DNA | 2.0 | 2.0 |
PCR of Toggle 1
Contents | ul | Phire | 10 |
---|---|
H2O | 8 |
5 uM F Primer | 1.5 |
5 uM R Primer | 1.5 |
100pg/ul Construct | 1.0 |
- Ran other trials with 2.5, 3.5, 4.5 and 6.5 ul of Forward and Reverse Primers
- Water ul was adjusted to compensate: 6, 4, 2 and 0 ul respectively
Phase | Temperature (°C) | Time (sec) |
---|---|---|
Initial Denaturation | 98 | 30 |
Denaturation | 98 | 5 |
Annealing | 68 | 5 |
Extension | 72 | 45 |
Final Extension | 72 | 60 |
PCR of Toggle 2
Contents | ul | Phire | 10 |
---|---|
H2O | 8 |
5 uM F Primer | 1.5 |
5 uM R Primer | 1.5 |
100pg/ul Construct | 1.0 |
- Ran other trials with 2.5, 3.5, 4.5 and 6.5 ul of Forward and Reverse Primers
- Water ul was adjusted to compensate: 6, 4, 2 and 0 ul respectively
Phase | Temperature (°C) | Time (sec) |
---|---|---|
Initial Denaturation | 98 | 30 |
Denaturation | 98 | 5 |
Annealing | 68 | 5 |
Extension | 72 | 45 |
Final Extension | 72 | 60 |
Digests of Toggle parts
Toggle 1 Digest
Content | Experimental ul | Control ul |
---|---|---|
H2O | 24 | 25 |
Cutsmart | 5.0 | 5.0 |
SpeI | 1.0 | X |
DNA | 25.0 | 25.0 |
Toggle 1 Digest
Content | Experimental ul | Control ul |
---|---|---|
H2O | 24 | 25 |
Cutsmart | 5.0 | 5.0 |
XbaI | 1.0 | X |
DNA | 25.0 | 25.0 |
Linearized pSB1C7 Digest
Content | Experimental ul | Control ul |
---|---|---|
H2O | 16.5 | 18.5 |
Cutsmart | 2.5 | 2.5 |
SpeI-HF | 1.0 | X |
XbaI | 1.0 | X |
DNA | 4.0 | 4.0 |
Ligation of Toggle parts
Content | ul |
---|---|
10X T4 Buffer | 2.0 |
Toggle 1 | 5.0 |
Toggle 2 | 10.0 |
H2O | 2.0 |
Ligase | 1.0 |