Coenen Lab Protocols

Below are protocols commonly used by students in my lab (PDFs).

If you have a nifty protocol you think would be useful to other students, I'd be happy to add it here.

Plant Culture

Plating tobacco seeds.    A basic protocol for growing Nicotiana plumbaginifolia seedlings in sterile culture.

Plating fern spores.     How to sow Ceratopteris spores for observation of gametophytes in sterile culture.

Molecular Biology

Ten Minute Miniprep.    A quick-and-easy protocol for preparing small amounts of plasmid DNA from E. coli. DNA prepared by this method is clean enough for restriction digests and PCR screening but should not be used for sequencing applications.

Plasmid Midiprep.   To prepare highly pure plasmid DNA using the Quiagen Midiprep Kit.

Agrobacterium Miniprep.    A quick procedure for purifying small amouts of plasmid DNA from Agrobacterium and other slimy bacteria. DNA is sufficiently pure for restriction analysis.

E. coli Transformation.   To introduce a plasmid into chemo-competent bacteria.

Restriction Digest.   Digesting plasmid DNA with restriction endonucleases.

DNA Analysis in Agarose Gels.   How to pour, load, and run agarose gels for analysis of restriction digests.

Gel Documentation.   How to take a picture of your DNA gel using the digital camera.

Media & Solutions

YEP.    Medium for growing Agrobacterium tumefaciens.

LB.    Medium for growing Escherichia coli.

SOC.     Medium for transformation of E. coli cells.

Equipment Use

Autoclave.    Basic do's and dont's of autoclave operation.

Koehler Illumination.   How to adjust your microscope for optimal image resolution.

Microscopy Primer. A website with a wealth of information on a wide variety of microscopic techniques.

pH meter.    In case you forgot / don't have your plant physiology lab manual handy: here's how to get most of the pH meters in the department to work well for you.

Pixera Digital Camera.   How to take pictures of small seedlings on agar plates or under the microscope.

Speed Vac.   How to vacuum-infiltrate samples / dry DNA pellets without getting pump oil sucked into the vacuum chamber.

Milton-Roy Spectrophotometer.   How to measure the absorbance of samples using the Milton Roy Spectronic 601.

Beckman Spectrophotometer. 

Miscellaneous

Chlorophyll & Anthocyanins.   A basic procedure for extracting and measuring chlorophyll and/or anthocyanin content of plant samples.

Paraplast Embedding of soft plant tissues for sectioning with the microtome.