*3.2 (from the 6 clones that were selected on 29/06/16)
*3.2 (from the 6 clones that were selected on 29/06/16)
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The same digestion protocol as on {{Font color||yellow|???}} was followed increasing the DNA quantity (10µL DNA, 7µL water, 2µL Red buffer, 0.5µL EcoR1, 0.5µL HindIII). Digestion products were migrated on agarose gel.
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{| class="wikitable"
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The DNA concentration was still insufficient meaning the extraction step was not efficient. New extraction will be performed on clones obtained after the transformation.
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|-
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! Component
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! Volume (µL)
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|-
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| Plasmid
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| 10
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|-
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| Red buffer 10x
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| 2
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|-
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| Water
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| 7
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|-
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| EcoRI enzyme
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| 0.5
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|-
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| HindIII enzyme
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| 0.5
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|}
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The mix was incubated at 37°C for 1 hour.
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An electrophoresis was done (0.8% of agarose).
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{| class="wikitable"
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|-
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! Component
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! Volume (µL)
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|-
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| Digested DNA
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| 20
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|-
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| Loading buffer
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| 3.3
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|}
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The DNA concentration was still insufficient meaning the extraction step was not efficient. Colony PCR
