NYMU Taipei/Protocols2

From 2007.igem.org

Revision as of 05:28, 21 October 2007

Wet-Lab Protocols

• Protocol online
• PCR (7/20)
  • RT-PCR
  • Q-PCR
• Quantitation of  Nucleic acid concentration
  • [Wikipedia]
  • [Pierce]
  • [India]
  • [Kreatech Biotechnology] 
• <span lang="EN-US" style="font-size: 9.5pt; font-family: Arial [Setting Up a Restriction Endonuclease Reaction]</span>
• DNA Analysis through Gel Electrophoresis
• Ligate blunt or cohesive ends in 5 minutes at room temperature
  • [Quick Ligation Kit (NEB)]
  • [Fast Ligation Kit (UBI)]
  • [Fast-Link DNA Ligation Kit (EPICENTRE Biotechnologies)]
  • [Rapid Ligation Kit (Fermentas)]
  • All of these are just normal ligase buffers with PEG8000 at a 10-12% concentration. The NEB catalogue gives the original reference. 
    Use of crowding agents like PEG allow 5-minute sticky end ligations.
• Vector construction
  • Vector sequences
    • [VectorDB - Molecular Biology Vector Sequence Database]
    • [The cloning vector collection]
    • [New England Biolabs (NEB) DNA Sequences and Maps]
    • [Promega Cloning Vectors]
    • [Novagen Vector Maps]
  • [PlasMapper Version 2.0 - automatically generates and annotates plasmid maps]
• Transformation
• Production of heterologous proteins in Escherichia coli
• [Antibiotics]
• [Part fabrication] ([Enzyme selection for BioBricks digest]!!)
• [BioBricks construction tutorial]
• [Fusion protein]
• Glucose assay
  • [Blood sugar (from Wikipedia)]
  • [Glucose Measurement for Cell Culture]
  • [Glucose Assay Kits from SigmaAldrich]
  • [Glucose Assay Kit from QuantiChrom]
  • [Biochemical glucose kit from rpbiotech]
  • [Glucose Meter from AnilinePharma]
• Mammalian Gene Collection (MGC) Full-length cDNA clones
  • [at National Cancer Institute of NIH]
  • [at Genome Research Center of NYMU]
  • [NCBI MGC retrieval]

Dry-Lab Protocols

• [serial cloner]
• Protein Fusion Design
  • rcsC-envZ cimaera design
  • Literature review
  • Computational analysis
• Twin-Arginine Translocation Pathway Signal Peptide
  • Lee et al., 2006. The Bacterial Twin-Arginine Translocation Pathway. Annu. Rev. Microbiol., 60, p373-395
    • Properties of TAT Signal Peptides
    • [A link to a list of Tat signal sequences provided by Tracy Palmer]
    • 29 putative signal peptides 
    • Selected TAT signal peptides
    • Signal peptides and insulin A, B chain
    • Signal peptides and IDE coding sequence

General references

• [New England Biolab (NEB) technical references]

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