SlrR was ordered from IDT with prefix and suffix. PCR was performed with the prefix and suffix primers.
Prefix primer:
CAGGCAGTTGAATTCGCGGCCGCTTCTAGA
Suffix primer:
CTTGAGCTCCTGCAGCGGCCGCTACTAGTA
The following thermocycle was observed.
Template:Team:Groningen/CONTENT/EXPERIMENTS/slrR PCR (sl1)
00:00, 28 July 2015 - 00:00, 28 July 2015
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Step
Temperature
Time
1
Initiation
98 °C
5 min
2
Denaturation
98 °C
10 sec
Annealing
55 °C
30 sec
Extension
72 °C
90 sec
3
Final extension
72 °C
5 min
Thermocycle.
step 2 was repeated 35 times. PCR product was the right fragment and was purified with a PCR purification kit.