Difference between revisions of "Davidson Missouri W/Davidson Protocols"
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| − | '''General Lab Information''' | + | '''A. General Lab Information''' |
# [http://www.bio.davidson.edu/courses/molbio/labnotebook.html How to Keep a Lab Notebook] | # [http://www.bio.davidson.edu/courses/molbio/labnotebook.html How to Keep a Lab Notebook] | ||
# [http://www.bio.davidson.edu/courses/Molbio/Protocols/reagents.html Common molecular reagents] | # [http://www.bio.davidson.edu/courses/Molbio/Protocols/reagents.html Common molecular reagents] | ||
| Line 9: | Line 9: | ||
| − | '''Gel Electrophoresis and Purification''' | + | '''B. Gel Electrophoresis and Purification''' |
# [http://www.bio.davidson.edu/courses/Molbio/Protocols/pourgel.html Pouring an agarose gel] | # [http://www.bio.davidson.edu/courses/Molbio/Protocols/pourgel.html Pouring an agarose gel] | ||
# [http://www.bio.davidson.edu/courses/Molbio/Protocols/molwt.html Calculate MWs] | # [http://www.bio.davidson.edu/courses/Molbio/Protocols/molwt.html Calculate MWs] | ||
| Line 18: | Line 18: | ||
# [http://www.bio.davidson.edu/courses/Molbio/Protocols/ligation.html Ligation Protocol] | # [http://www.bio.davidson.edu/courses/Molbio/Protocols/ligation.html Ligation Protocol] | ||
| − | '''Digestion, Ligation, Transformation''' | + | '''C. Digestion, Ligation, Transformation''' |
# [http://www.bio.davidson.edu/courses/Molbio/Protocols/digestion.html Digest DNA with restriction enzymes] | # [http://www.bio.davidson.edu/courses/Molbio/Protocols/digestion.html Digest DNA with restriction enzymes] | ||
# [[Davidson Missouri W/Double Digest Guide| Double Digest Guide]] | # [[Davidson Missouri W/Double Digest Guide| Double Digest Guide]] | ||
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# [[TSS Competent Cells|TSS Competent Cell Transformation]] | # [[TSS Competent Cells|TSS Competent Cell Transformation]] | ||
| − | '''Minipreps''' | + | '''D. Minipreps''' |
# [http://www.bio.davidson.edu/courses/Molbio/Protocols/MiniPrep_list.html Choices for Mini-Preps: Promega vs. Zyppy] | # [http://www.bio.davidson.edu/courses/Molbio/Protocols/MiniPrep_list.html Choices for Mini-Preps: Promega vs. Zyppy] | ||
# [http://www.bio.davidson.edu/courses/Molbio/Protocols/miniprepPrmega.html Promega miniprep] | # [http://www.bio.davidson.edu/courses/Molbio/Protocols/miniprepPrmega.html Promega miniprep] | ||
# [http://www.bio.davidson.edu/courses/Molbio/Protocols/Zippy_MiniPrep.html Zippy Miniprep] | # [http://www.bio.davidson.edu/courses/Molbio/Protocols/Zippy_MiniPrep.html Zippy Miniprep] | ||
| − | '''Making New Parts and PCR''' | + | '''E. Making New Parts and PCR''' |
# [http://www.bio.davidson.edu/courses/Molbio/Protocols/anneal_oligos.html Building dsDNA with Oligos] | # [http://www.bio.davidson.edu/courses/Molbio/Protocols/anneal_oligos.html Building dsDNA with Oligos] | ||
# [http://www.bio.davidson.edu/courses/Molbio/Protocols/pcr.html Setting up PCR mixtures] | # [http://www.bio.davidson.edu/courses/Molbio/Protocols/pcr.html Setting up PCR mixtures] | ||
| Line 40: | Line 40: | ||
# [http://www.bio.davidson.edu/courses/Molbio/Protocols/ColonyPCR_Screening.html Colony PCR to Screen for Successful Ligations] | # [http://www.bio.davidson.edu/courses/Molbio/Protocols/ColonyPCR_Screening.html Colony PCR to Screen for Successful Ligations] | ||
| − | '''Expression of Phenotypes''' | + | '''F. Expression of Phenotypes''' |
# [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC106306/pdf/am002240.pdf Using degradation tags on proteins such as GFP] | # [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC106306/pdf/am002240.pdf Using degradation tags on proteins such as GFP] | ||
# [[Genomic Insertion Protocol|Genomic Insertion Protocol]] | # [[Genomic Insertion Protocol|Genomic Insertion Protocol]] | ||
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# When inducing with Arabinose, use "2 µL of stock" (10% w/v L-Arabinose) "to every 1 mL" of LB or other liquid. | # When inducing with Arabinose, use "2 µL of stock" (10% w/v L-Arabinose) "to every 1 mL" of LB or other liquid. | ||
| − | '''Computer Tools We Use''' | + | '''G. Computer Tools We Use''' |
# [http://gcat.davidson.edu/iGEM08/tools.html All Sites In One Place] | # [http://gcat.davidson.edu/iGEM08/tools.html All Sites In One Place] | ||
# [http://gcat.davidson.edu/iGEM08/gelwebsite/gelwebsite.html Optimize your Gel] | # [http://gcat.davidson.edu/iGEM08/gelwebsite/gelwebsite.html Optimize your Gel] | ||
Revision as of 21:22, 13 March 2010
A. General Lab Information
- How to Keep a Lab Notebook
- Common molecular reagents
- Standard Assembly
- BioBrick Ends
- Compatibility of Plasmids
- Ethanol Precipitate DNA (short protocol)
- glycerolstocks How to Make Glycerol Stocks of Bacteria
B. Gel Electrophoresis and Purification
- Pouring an agarose gel
- Calculate MWs
- 1kb MW markers
- Qiagen QIAquick Gel Purification
- Qiagen QIAquick Column Regeneration Protocol
- ElectroElute Gel Purification
- Ligation Protocol
C. Digestion, Ligation, Transformation
- Digest DNA with restriction enzymes
- Double Digest Guide
- Shrimp Alkaline Phosphatase
- Choices for Transformation: Heat Shock vs. Zyppy
- Heat Shock Transformation OR Short version of Heat Shock
- Zippy Transformation
- TSS Competent Cell Transformation
D. Minipreps
E. Making New Parts and PCR
- Building dsDNA with Oligos
- Setting up PCR mixtures
- PCR and Mg2+ concentration
- Making dsDNA Using Primer Dimers
- Clean and Concentrate DNA (after PCR, before digestion)
- Colony PCR to Screen for Successful Ligations
F. Expression of Phenotypes
- Using degradation tags on proteins such as GFP
- Genomic Insertion Protocol
- When inducing with IPTG, use 3 µL of stock (0.2 g/mL = 20% w/v) to every 1 mL of LB or other liquid.
- When inducing with Arabinose, use "2 µL of stock" (10% w/v L-Arabinose) "to every 1 mL" of LB or other liquid.
G. Computer Tools We Use
- All Sites In One Place
- Optimize your Gel
- Gene Splitting Web Site
- PCR Primers w/ BioBricks
- Promega Tm Calculator
- List of auto-inducers and their catalog numbers
- Sequencing at Agencourt Bioscience
- Sequencing at CUGI
- Analyzing Sequences with aPe
- Lance-olator Oligos for dsDNA assembly
- Access the GCAT-alog of Davidson and MWSU DNA Freezer Stocks
- Using Apes (A Plasmid Editor)
