User:Mike Barnkob

Information

• Mike Barnkob
• Website: www.mikebarnkob.dk.

Protocols

Cloning

• Basics
  • Solutions
  • Primer design
  • Gels
  • Isolating mRNA from spleen
  • Creating cDNA from mRNA
  • PCR
• CRISPR / Cas9
  • Designing sgRNA oligos
  • Cloning oligos into backbone
  • Transforming bacteria with construct
  • Screening for inserts
• Lentiviral work
  • Lentiviral production
  • Infecting target cells

Bacterial work

• Prepare antibiotics
• LB agar plates
• Streaking plates
• Inoculate overnight liquid culture
• Glycerol stock
• Isolate plasmid DNA
• Diagnostic Restriction Digest

Immunology

• Flow cytometry
  • General notes
  • Generic protocol
  • Preparing beads
• CFSE staining
• T cell enrichment from tumours
• Making tetramers

Tissue culture work

• Media
• Thawing cells
• Freezing down cells
• Split cells
• Count cells
• Harvest splenocytes
• Harvest BMDCs
• In vitro expansion of T cells
• Soft agar assay

Visualization

• Long-time co-culture and visualization with live-dead marker
• Immunofluorescence on cell lines

Basics

• Formulars

Projects

I work on:

• T cell and cancer cell interactions
• Genetic manipulation of T cells

Past:

Liquid handling robot. An attempt to create a small liquid handling robot to do simple experiments with.

Bacto Bandage. 2009 SDU Denmark iGEM team. Our goal was to create an E. coli strain, which inhibits S. aureus biofilm formation in wounds by producing RNA III-inhibiting-peptide (RIP).