Versions Compared

Key

  • This line was added.
  • This line was removed.
  • Formatting was changed.

Antibody Entry Vector

Take G Blocks Order G blocks "mIgM heavy chain against beta amyloid" and "mIgM kappa chain against beta amyloid" that are flanked with complementary sequences to Nelson's primer with Q1 and QX Qx sites

PCR amplify G blocks with respective Nelson's primers

Golden Gate G blocks into pGDONR Vectorgate G blocks into a pDONR vector creating pENTR L1-mIgMheavy-L2 and pENTR L1-mIgMkappa-L2

Transform bacteria and plate on lactose+ media

Miniprep out pENTR L1-mIgMheavy-L2 and pENTR L1-mIgMkappa-L2 vectors 

LR pENTR L1-mIgMheavy-L2 and pENTR L1-mIgMkappa-L2 vectors each with pENTR L4-Hef1a-R1 to create pEXPR hef1a-mIgMheavy and pEXPR hef1a-mIgMkappa for expression of the antibody

Transform bacteria again?