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 |  |  |  |  |  |  | Transfection Request Form
 |  |  |  |  |  |  | University of Maryland, Baltimore    School of Medicine Transgenic Core Laboratory   Howard Hall 581    706-0453
 |  |  |  |  |  | Request Date
 |  |  |  |  |  | PI Name |  |  |  |  | Campus Address
 |  |  |  |  |  | Greenebaum Cancer Center Member? |  |  |  |  | (Office)
 |  |  |  |  |  |  | YES | NO |  |  | (Lab)
 |  |  |  |  |  | Department
 |  |  |  |  |  | Phone |  |  |  |  | email
 |  |  |  |  |  | Lab Contact |  |  | Phone |  |  |  | email
 |  |  |  |  |  | Chartstring to be charged
 |  |  |  |  |  | Animal Space Location: |  |  |  |  |  | ARF# |  | Exp. Date
 |  |  |  |  |  |  |  |  |  |  |  | Please include a copy of your protocol approval letter
 |  |  |  |  |  | Please provide 500 ug of linearized plasmid DNA for each project as ETOH ppt. DNA under 70% ETOH wash
 |  |  |  |  |  | Construct Name:
 |  |  |  |  |  |  |  | Please provide 1) a linear map of your construct 2) gel picture of linearized plasmid showing complete digestion
 |  |  |  |  |  | Positive selection cassette |  |  |  |  |  |  | promoter
 |  |  |  |  |  | Negative selection cassette |  |  |  |  |  |  | promoter
 |  |  |  |  |  | Total DNA provided (ug) |  |  |  |  | A260:280 |  | [NAME] (kb)
 |  |  |  |  |  | Source of ARM DNA (strain) |  |  |  | ARM length (kb) |  | 5'ARM |  | 3'ARM
 |  |  |  |  |  | Cells will be provided from 96-well plate cultures of ES clones that survive slection 3-4 weeks after electroporation. DNA can be
 |  |  |  |  |  | provided in place of cells at additional cost. Please report screening results promptly (2 weeks) to the Transgenic Lab at 6-0453.
 |  |  |  |  |  | No. of wells requested (circle one): |  |  | 192 |  | 288 |  | 384
 |  |  |  |  |  | Screening Assay: |  | PCR | Southern |  | Provide a gel photo (PCR) of the screening assay indicating 1) positive control,
 |  |  |  |  |  | (circle one) |  |  |  |  | 2) negative control and 3) demonstration of 1-copy gene per genome sensitivity
 |  |  |  |  |  | NOTE: Your screening assay must be able to distinguish homologous recombinants from non-homologous recombinants. Additionally the
 |  |  |  |  |  | screening assay must be able to detect a single gene copy per genome. The Transgenics Lab can give you guidelines to reach these goals.
 |  |  |  |  |  | A screening panel will be submitted to verify assay specificity and sensitivity, results must be received  prior to transfection of ES cells.
 |  |  |  |  |  | Do you require the Transgenic Lab to prep DNA from ES cell clones? |  |  |  |  |  | YES | NO
 |  |  |  |  |  | Please provide a project description as it pertains to the use of knockout animals:
 |  |  |  |  |  | Has this construct been used to generate knockout animals previously? |  |  |  |  |  |  | YES | NO
 |  |  |  |  |  | Are phenotypic problems expected (i.e. [NAME])? |  |  |  |  |  |  | YES | NO
 |  |  |  |  |  |  |  |  |  |  |  | if yes, explain
 |  |  |  |  |  | PI Signature: |  |  |  |  |  |  |  | Date
 |  |  |  |  |  | Transgenic Lab Approval: |  |  |  |  |  |  |  | Date
 |  |  |  |  |  | Return this completed form to: Transgenic Core Facility, [NAME] (FAX [PHONE])
 |  |  |  |  |  | This form and all requested information and supporting data must be received prior to project initiation