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[[File:2746 2770 2779 PCR.JPG|RTENOTITLE]] PCR from genomic DNA using superhifi II enzyme mix 6/6/2020 [[File:2763 2772 2773 2779 PCR.JPG|RTENOTITLE]] PCR again from genomic DNA [[File:2763 2779 2770 PCR.JPG|RTENOTITLE]] [[File:2770 2779 PCR.JPG|RTENOTITLE]] [[File:2773 2779 digestion.JPG|RTENOTITLE]] [[File:2772 2746 2779 2778.JPG|RTENOTITLE]] [[File:2779 pJet XhoI XbaI.JPG|RTENOTITLE]] 20200616 NdeI+HindIII, buffer2.1 ,37oC, 1hour [[File:2746 2763 2773 2779 pJet digestion.JPG|RTENOTITLE]] == 2779 gibson == 20200617 to get the vector from #483 digest with NdeI+HindIII in Buffer 2, 37oC , 40min [[File:2779 gibson vector.JPG|RTENOTITLE]] == Protein Expression == [[File:2779|RTENOTITLE]][[File:2779 protein expression.jpg|RTENOTITLE]] {| border="1" cellspacing="1" cellpadding="1" style="width:500px;" |- | Lane | Name | Desc |- | 1 | N protein 2 ul | <br/> |- | 2 | N protein 5 ul | <br/> |- | 3 | S protein 5 ul | BEI |- | 4 | RBD 5 ul | BEI |- | 5 | 2779 Turner 37 oC 2 hours, 7.5 ml, 0.2% rhamnose | <br/> |- | 6 | 2779 NEB 37 oC 2 hours, 7.5 ml, 0.2% rhamnose | <br/> |- | 7 | 2746 1mM IPTG | <br/> |- | 8 | 2723 | <br/> |- | 9 | 2779 NEB 25 oC o/n 7.5 ml, 0.2% rhamnose | cell dead |- | 10 | 2779 NEB 25 oC o/n 7.5 ml, 0.2% rhamnose | cell dead |} [[Category:Pages with broken file links]] == http://18.206.161.119/eln/images/6/66/2779_protein_exp_screening2.jpg == [[Category:Pages with broken file links]] [[Category:Cloning]]
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