Haynes Lab:Notebook/ASU iGEM

ASU iGEM 2012

Project Description/Abstract

Team Chimeric Reporter:

• Hyder Hussain
• Abhinav Markus
• Ryan Muller
• Ethan Ward

Team Cell Surface Reporter:

• Nisarg Patel
• Ryan Muller
• Rohit Rajan
• Ellen Qin
• Melinda Jenner
• Joe Barth

BioBricks

Magainin Reporter

• Magainin
• Magainin + 2xGS Linker + His-Tag
• Bgal fragment (a-1)
• Bgal fragment (a-4)
• Bgal fragment (w)
• Bgal fragment (1-w)
• Magainin + 2aa GS Linker + His-Tag + 10aa GS Linker + Bgal fragment (a-1)
• Magainin + 2aa GS Linker + His-Tag + 10aa GS Linker + Bgal fragment (a-4)
• Magainin + 2aa GS Linker + His-Tag + 10aa GS Linker + Bgal fragment (w)
• Magainin + 2aa GS Linker + His-Tag + 10aa GS Linker + Bgal fragment (1-w)
• His-Tag + 10aa GS Linker + Bgal fragment (a-1)
• His-Tag + 10aa GS Linker + Bgal fragment (a-4)
• 13. His-Tag + 10aa GS Linker + Bgal fragment (w)
• 14. His-Tag + 10aa GS Linker + Bgal fragment (1-w)

Notes

6-15-12

• Competent cell prep

• Centrifuged falcon test tubes containing liquid colonies
• Resuspended in CaCl₂ buffer solution and incubated for 15 mins
• Centrifuged and resuspended in CaCl₂ glycerol buffer solution
• Chilled overnight

6-16-12

• Competent cell prep

• Aliquotted 200uL into test tubes
• Stored in -80C

6-17-12

• Streak plated prepared competent cells on LB no amp plate

• Colonies observed

6-19-12

• Transformation (LSE)

• Transformed DNA:

• T7 promoter BBa_I712074
• Constitutive promoter BBa_J23102

• Cells: DH5 alpha (donated)
• Protocol from: http://tools.invitrogen.com/content/sfs/manuals/subcloningefficiencydh5alpha_man.pdf (30 minute transformation)
• Controls: puc19, no DNA
• Plated 2 copies of each (100 ul, 250 ul) on LB amp plates.

• Made 50 LB Amp plates.