How To Make A MEGA-plate

Overview

This page provides step-by-step instructions for the MEGA-Plate experiment for evolving and tracking microbial evolution on antibiotic gradients {ref}.\n The MEGA-plate was developed by Michael Baym, Tami Liemberman, Eric Kelsic, Remy Chait, Rotem Gross, Idan Yelin, Roy Kishomy Kishony Lab at Harvard Medical School and Technion - Israel Institute of Technology

Materials

For a plate of 62cm on 82cm:

• Polycarbonate sheet 62cm X 82cm X 0.6cm (compeny,ID)
• Plexiglas sheet 30cm X 60cm X 0.3cm (compeny,ID)
• Dichloromethane (compeny,ID)

• Syringe (compeny,ID)
• Needle (compeny,ID)
• 9 X 2L Erlenmeyer flask (compeny,ID)

• Agar (http://www.bd.com/ds/productCenter/214010.asp#)
• LB Broth (http://www.bd.com/ds/productCenter/240220.asp)

Procedure

1. Plan the design of the plate.

1. Use laser cutter to cut the plexiglas sheet to your design:
   1. 8 pieces of 40cm X 1.27cm
   2. 4 pieces of 40cm X 2cm
   3. 2 pieces of 60cm X 2cm

1. Glue the pieces like the plan design.
2. Check for leaking.
3. Use blith

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

1. List troubleshooting tips here.
2. You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
3. Anecdotal observations that might be of use to others can also be posted here.

Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.

References

Relevant papers and books

1. Goldbeter, A and Koshland, DE (1981) - Proc Natl Acad Sci U S A 78(11) 6840-4 PMID 6947258
2. Jacob, F and Monod, J J (1961) - Mol Biol 3(3) 318-56 PMID 13718526
3. Ptashne, M (2004) Genetic Switch: Phage Lambda Revisited - Cold Spring Harbor Laboratory Press ISBN 0879697164

Contact

This page was developed by Michael Baym and Rotem Gross from the Kishony lab. For questions, please contact: Michael Baym (mbaym@gmail.com) Rotem Gross () Roy Kishony (rkishony@technion.ac.il)

or instead, discuss this protocol.