Team:DTU-Denmark/Notebook/8 September 2013
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=lab 208= | =lab 208= | ||
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==Who was in the lab== | ==Who was in the lab== | ||
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===Colony PCR to verify Nir in pZA21::ara-tight=== | ===Colony PCR to verify Nir in pZA21::ara-tight=== | ||
- | == | + | Colony PCR on the Nir transformants, testing for NirH (primers 45a, 45b, annealing 59C), NirM+S (primers 44, 11a annealing 64C) and NirN (primers 43, 11b annealing 68C). |
- | + | ||
+ | program: | ||
+ | {| class="wikitable" style="text-align: right" | ||
+ | ! temperature !! time !! cycles | ||
+ | |- | ||
+ | | 98C || 10:00 || - | ||
+ | |- | ||
+ | | 98C || 0:10 || 36 | ||
+ | |- | ||
+ | | 59C/64C/68C || 0:30 || 36 | ||
+ | |- | ||
+ | | 72C || 2:00 || 36 | ||
+ | |- | ||
+ | | 72C || 5:00 || - | ||
+ | |- | ||
+ | | 10C || hold || - | ||
+ | |- | ||
+ | |} | ||
+ | |||
+ | Colony PCR to verify HAO insert. | ||
+ | |||
+ | Primers: 28a, 28b | ||
+ | |||
+ | temperature: 59C | ||
+ | |||
Latest revision as of 12:12, 4 October 2013
8 September
Contents |
lab 208
Who was in the lab
Henrike, Kristian, Julia
Procedure
Colony PCR to verify Nir in pZA21::ara-tight
Colony PCR on the Nir transformants, testing for NirH (primers 45a, 45b, annealing 59C), NirM+S (primers 44, 11a annealing 64C) and NirN (primers 43, 11b annealing 68C).
program:
temperature | time | cycles |
---|---|---|
98C | 10:00 | - |
98C | 0:10 | 36 |
59C/64C/68C | 0:30 | 36 |
72C | 2:00 | 36 |
72C | 5:00 | - |
10C | hold | - |
Colony PCR to verify HAO insert.
Primers: 28a, 28b
temperature: 59C
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