Materials
Name | Company | Catalog Number | Comments |
Agar | Prolabo | 20768.292 | for NGM (nematode growth medium) preparation (6,7) |
NaCl | Chimie-Plus | 40045 | for NGM preparation |
Bacto-peptone | BD | 211820 | for NGM preparation |
KH2PO4 | Roth | 3904-1 | for NGM preparation |
K2HPO4 | Prolabo | 26930.293 | for NGM preparation |
MgSO4 | Roth | P027.2 | for NGM preparation |
CaCl2 | Prolabo | 22317.297 | for NGM preparation |
Cholesterol | Rectapur | for NGM preparation | |
LB-broth | Sigma | L3022 | for culture of OP50 bacteria |
EQUIPMENT | |||
U.V. cross-linker | Fischer Scientific | Wavelength 254 nm | |
Microscope SteREO Discovery V8 | Carl Zeiss, Inc. | ||
Petri dishes, 60 mm | CML | BPS55E6 | |
Platinium wire 0.25 mm | Alfa Aesar | 7440-06-4 | To make a pick for worms (7) |
References
- Berkowitz, L. A., Knight, A. L., Caldwell, G. A., Caldwell, K. A. Generation of stable transgenic C. elegans using microinjection. J. Vis. Exp. (18), e833 (2008).
- Mello, C., Kramer, J. M., Stinchlomb, D., Ambros, V. Efficient gene transfer in C.elegans: extrachromosomal maintenance and integration of transforming sequences. EMBO J. 10 (12), 3959-3970 (1991).
- Transformation and microinjection. WormBook. Evans, T. C. , The C. elegans Research Community, WormBook. (2006).
- Hobert, O. PCR Fusion-Based Approach to Create Reporter Gene Constructs for Expression Analysis in Transgenic C. elegans. BioTechniques. 32, 728-730 (2002).
- Stinchcomb, D. T., Shaw, J. E., Carr, S. H., Hirsh, D. Extrachromosomal DNA transformation of Caenorhabditis elegans. Mol. Cell. Biol. 5, 3484-3496 (1985).
- Brenner, S.
The genetics of Caenorhabditis elegans. Genetics. 77, 71-94 (1974). - Chaudhuri, J., Parihar, M., Pires-daSilva, A. An Introduction to Worm Lab: from Culturing Worms to Mutagenesis. J. Vis. Exp. (47), e2293 (2011).
- Merritt, C., Rasoloson, D., Gallo, C., Voronina, E., Ko, D., Seydoux, G. A modular vector kit for transgenesis in C. elegans. International Worm Meeting. , (2007).
- Chalfie, M., Tu, Y., Euskirchen, G., Ward, W., Green Prasher, D. Fluorescent protein as a marker for gene expression. Science. 263, 802-805 (1994).
- Fleenor, J., Fire, A. Discosoma sp. Red Fluorescent Protein (dsRED) as a reporter in C. elegans. Worm Breeder's Gazette. 16 (3), 16 (2000).
- Lecroisey, C., et al. ZYX-1, the unique zyxin protein of Caenorhabditis elegans, is involved in dystrophin-dependent muscle degeneration. Mol. Biol. Cell. , (2013).
- Praitis, V., Casey, E., Collar, D., Austin, J. Creation of low-copy integrated transgenic lines in Caenorhabditis elegans. Genetics. 157 (3), 1217-1226 (2001).
- Frøkjaer-Jensen, C., et al. Single-copy insertion of transgenes in Caenorhabditis elegans. Nat. Genet. 40 (11), 1375-1383 (2008).
- Dickinson, D. J., Ward, J. D., Reiner, D. J., Goldstein, B. Engineering the Caenorhabditis elegans genome using Cas9-triggered homologous recombination. Nat. Methods. 10 (10), 1028-1034 (2013).