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iGEM 2010
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Overview Toggle Phage Mammalian Summary Acknowledgements
Materials & Methods Biosafety Journal Club
Bacterial Construction Protocol
Bacterial Experimental Protocol
Phage western blot
Mammalian Cell Lines
Microfluidic stress
Materials
biobrick construction |
Miniprep To extract DNA from cells, we used the QIAprep Spin Miniprep Kit according to protocol. DNA Digestion For both restriction mapping and for digesting a plasmid to insert or extract a biobrick, the appropriate buffer was added 1:10 to a DNA sample and 10 units of the appropriate enzyme(s) were used per 1ug of DNA. The reactions were allowed to run at the temperature optimal for the enzyme for 1 hour, the enzymes were denatured at 65°C for 20 minutes, and then the reactions were held at 16°C. Antarctic Phosphatase To prevent recircularization of digested plasmids, we used NEB's Antarctic Phosphatase according to protocol. Gel Electrophoresis To view lengths of DNA, Orange G 6x was added 1:3 to 300ng miniprepped DNA and run against 5uL of Hyperladder I in the lanes of 1% UltraPure agarose gels in TAE buffer at 120V for 45 minutes. DNA Gel Extraction To extract the DNA from the agarose gel, the QIAquick Gel Extraction kit was used according to protocol. |
