Nematocida sp. 1 strain线虫 BioVector NTCC质粒载体菌种细胞基因保藏中心

Nematocida sp. 1 strain线虫 BioVector NTCC质粒载体菌种细胞基因保藏中心
Strain Designation: ERTm6
Depositor: ER Troemel
Isolation:
Wild­caught Caenorhabditis briggsae, Cape Verde islands
Frozen ampules packed in dry ice should either be thawed immediately or stored in liquid nitrogen. If liquid
nitrogen storage facilities are not available, frozen ampoules may be stored at or below ­70°C for
approximately one week. Do not under any circumstance store frozen ampules at refrigerator
freezer temperatures (generally ­20°C). Storage of frozen material at this temperature will result in the
death of the culture.
1. To thaw a frozen ampule, place it in a 35°C water bath such that the lip of the ampule remains above the
water line. Thawing time is approximately 1 minute. Do not agitate the ampule. Do not leave ampule in water
bath after it is thawed.
2. When completely thawed, dilute the spore preparation by addition of 0.25 to 0.5 mL of a balanced saline
buffer solution such as ATCC medium 1323 Page's Balanced Saline, M9 buffer, or a Phosphate Buffered
Saline (PBS; ATCC cat. 30­2200).
3. Infect C. elegans nematodes by adding the dilute spore preparation onto an agar plate containing an
established C. elegans culture (stage L1 or L4/young adults). Seal the plate with parafilm and incubate
upright at 25°C. The worms become infected by simply ingesting the spores. Follow the protocol for
maintenance in­vivo.
Culture Maintenance
MAINTENANCE OF HOST C. ELEGANS CULTURE:
C. elegans nematodes may be maintained at 25°C on agar plates of ATCC medium 997, NG, or similar media
containing a lawn of E. coli bacteria. If desired, a uracil­requiring strain of E. coli (such as strain OP50­1,
available from the Caenorhabditis Genetics Center, University of Minnesota) may be used on a growth medium
containing limited uracil, thereby precluding bacterial overgrowth which may obscure view of the nematodes.
NG agar
Agar (Difco 214010) 17.0 g
NaCl 3.0 g
Bacto peptone (Difco 211677) 2.5 g
Cholesterol (5 mg/mL) 1.0 mL
CaCl
2 (1 M) 1.0 mL
MgSO4 (1 M) 1.0 mL
KH
2PO4 (1 M) 25.0 mL
Mix the first three reagents in 800 mL of distilled water and autoclave. After the mixture is cool, add the last
four reagents aseptically and adjust the volume of the medium to 1 L with sterile distilled water. See
www.atcc.org for ATCC medium formulations. For further information on cultivation of C. elegans nematades,
reference the following:
Brenner, S, 1974. Genetics 77: 71–94.
MAINTENANCE OF NEMATOCIDA CULTURE IN­VIVO:
Nematocida infection typically results in 50% host worms showing symptoms of infection at 2­3 d, and 50%
host worm mortality at 4­5d. The infection may be propagated by transfer of a few (<10) infected host worms
to a fresh plate culture of uninfected nematodes. Observe worms daily for symptoms of infection by
preparing wet mounts for phase microscopy at approximately 600x (DIC imaging recommended). Nematocida
infection manifests as distinct, granule­free regions within the C. elegans intestinal
【Supplier来源】BioVector NTCC Inc.
TEL：+86-010-53513060
【Website网址】 http://www.biovector.net