cDNA synthesis

• first strand cDNA synthesis with Ready-To-Go Beads (Amersham)
• first strand cDNA synthesis with SuperScript II
• double stranded cDNA synthesis with Time Saver Kit (Amersham)

Cell biology

• Cell culture basics
• Monocytes/Macrophages: enrichment of PBMCs with monocytes
• Monocytes/Macrophages: monocyte isolation using CD14 microbeads

RNA related methods

• RNA isolation with RNeasy mini kit

DNA related methods

• High Fidelity PCR
• Ligation of a fragment into a plasmid vector
• Plasmid miniprep with Qiagen buffers
• Restriction enzymes - overview and protocols
• Transformation of chemically competent E.coli

DNA methylation analysis

• Review
• DNA methylation analysis using restriction enzyme digestion
• DNA methylation analysis using bisulphite sequencing v.2 (2007)
• DNA methylation analysis using bisulphite sequencing
• DNA methylation analysis using Single Nucleotide Primer Extension (SNuPE)

Flow cytometry guides, protocols and software

Histology, Immunohistochemistry, FISH

• Fluorescent In Situ Hybridisation (FISH)
• HOPE tissue fixation
• Immunohistochemistry with Envision reagents

Immunology

• ELISA with DuoSet

Screenings

• PCR-based library screening method (PDF only!)

Mutagenesis

• Introducing mutations using PCR splicing

PCR

• Fibronectin EDA splicing
• PDH
• Quantitative PCR (qPCR, real-time PCR)

Protein methods

• Protein arginine methylation analysis
• Protein phosphorylation analysis

Transfection

• Transfection reagent selection
• Transfection normalizing using dot-blot hybridization