NYMU Taipei/Protocols2

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Revision as of 16:44, 25 October 2007

Wet-Lab Protocols

• [http://www.protocol-online.org/ Protocol online]
• PCR (7/20)
  • RT-PCR
  • Q-PCR
• Quantitation of  Nucleic acid concentration
  • http://en.wikipedia.org/wiki/Quantification_of_nucleic_acids Wikipedia
  • http://www.piercenet.com/Objects/View.cfm?type=Page&ID=0A0A7BB1-7582-4611-A116-6F583EC01666 Pierce
  • http://mcbl.iisc.ernet.in/Welcome to MCBL/Faculty/Parag/microarray workshop details/quantitation.html India
  • http://www.kreatech.com/pages/Focus-Areas/*-Hybridization-___-Labeling-protocols/Small-RNA-Labeling-protocol/ Kreatech Biotechnology 
• <span lang="EN-US" style="font-size: 9.5pt; font-family: Arial http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/setting_up_reaction.asp Setting Up a Restriction Endonuclease Reaction</span>
• DNA Analysis through Gel Electrophoresis
• Ligate blunt or cohesive ends in 5 minutes at room temperature
  • http://www.neb.com/nebecomm/products/productM2200.asp Quick Ligation Kit (NEB)
  • http://www.ubi.ca/cart/index.php/cPath/27_163_165 Fast Ligation Kit (UBI)
  • http://www.epibio.com/item.asp?ID=296 Fast-Link DNA Ligation Kit (EPICENTRE Biotechnologies)
  • http://www.fermentas.com/catalog/kits/kitrapidligation.htm 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
    • [[http://seq.yeastgenome.org/vectordb/ VectorDB - Molecular Biology Vector Sequence Database]]
    • [[http://www.shigen.nig.ac.jp/cvector/cvector.html The cloning vector collection]]
    • [[http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/dna_sequences_maps.asp New England Biolabs (NEB) DNA Sequences and Maps]]
    • [[http://www.promega.com/vectors/cloning_vectors.htm Promega Cloning Vectors]]
    • [[http://www.emdbiosciences.com/Products/BrowseProductsByCategory.asp?catid=1628 Novagen Vector Maps]]
  • [[http://wishart.biology.ualberta.ca/PlasMapper/ PlasMapper Version 2.0 - automatically generates and annotates plasmid maps]]
• Transformation
• Production of heterologous proteins in Escherichia coli
• [[http://openwetware.org/wiki/Endy:Preparing_Antibiotic_Stocks Antibiotics]]
• [[http://openwetware.org/wiki/Synthetic_Biology:BioBricks/Part_fabrication Part fabrication]] ([[http://openwetware.org/wiki/Enzyme_selection_for_BioBricks_digest Enzyme selection for BioBricks digest]]!!)
• [[http://openwetware.org/wiki/BioBricks_construction_tutorial BioBricks construction tutorial]]
• [[http://openwetware.org/wiki/Silver:_BB_Strategy Fusion protein]]
• Glucose assay
  • http://en.wikipedia.org/wiki/Blood_sugar Blood sugar (from Wikipedia)
  • [[http://www.genengnews.com/articles/chtitem.aspx?tid=1624&chid=3 Glucose Measurement for Cell Culture]]
  • [[http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Metabolomics/Product_Highlights/Enzymatic_Kits.html Glucose Assay Kits from SigmaAldrich]]
  • [[http://www.bioassaysys.com/Order.html Glucose Assay Kit from QuantiChrom]]
  • [[http://www.rpbiotech.com/Biochem_diag_kits/Glucose_Diagnostic_Kit.htm Biochemical glucose kit from rpbiotech]]
  • [[http://www.anilinepharma.com/products.php?catg=ds&id=1 Glucose Meter from AnilinePharma]]
• Mammalian Gene Collection (MGC) Full-length cDNA clones
  • [[http://mgc.nci.nih.gov/ at National Cancer Institute of NIH]]
  • [[http://genome.ym.edu.tw/libres/MGC_clones/index.htm at Genome Research Center of NYMU]]
  • [[http://www.ncbi.nlm.nih.gov/FLC/getmgc.cgi NCBI MGC retrieval]]

Dry-Lab Protocols

• http://serialbasics.free.fr/Serial_Cloner.html 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
    • http://www.jic.bbsrc.ac.uk/staff/tracy-palmer/signals.htm 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

• http://www.neb.com/nebecomm/tech_reference/default.asp New England Biolab (NEB) technical references

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