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Project:Public Biobrick: Difference between revisions
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=====Restriction digest===== | =====Restriction digest===== | ||
Required mass of DNA in each reaction is 250ng, so quantities of PSB1C3 (50.9ng/ul) and ANF (19.2ng/ul) are calculated with this in mind. EcoR1 and PST1 are the restriction enzymes. BSA prevents adhesion of the enzyme(s) to reaction tubes and pipette surfaces, and stabilizes some proteins during incubation. NEB2 is a buffer for the restriction enzymes. | Required mass of DNA (ie. plasmid or gene) in each reaction is 250ng, so quantities of PSB1C3 (50.9ng/ul conc.) and ANF (19.2ng/ul conc.) are calculated with this in mind. EcoR1 and PST1 are the restriction enzymes. BSA prevents adhesion of the enzyme(s) to reaction tubes and pipette surfaces, and stabilizes some proteins during incubation. NEB2 is a buffer for the restriction enzymes. | ||
'''Plasmid backbone''' | '''Plasmid backbone reaction''' | ||
(Total volume = 25ul) | |||
* PSB1C - 4.9ul | * PSB1C - 4.9ul | ||
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* NEB2 - 2.5ul | * NEB2 - 2.5ul | ||
'''ANF | '''ANF reaction'' | ||
(Total volume = 25ul) | |||
* ANF - 13ul | * ANF - 13ul | ||
| Line 147: | Line 147: | ||
* NEB2 - 2.5ul | * NEB2 - 2.5ul | ||
'''Control for ligation''' | '''Control for ligation reaction''' | ||
(Total volume = 25ul) | |||
* PSB1C - 4.9ul | * PSB1C - 4.9ul | ||
