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Nematocida parisii
Nematocida parisii
規(guī)格:
貨期:
編號:B221869
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱 Nematocida parisii
商品貨號 B221869
Strain Designations ERTm1
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Isolation
Wild-caught Caenorhabditis elegans isolated from a compost pit, Franconville, France
Product Format frozen
Storage Conditions Frozen: -70°C or colder for 1 week, vapor phase of liquid nitrogen for long-term storage
Type Strain no
Comments
Reference strain for genome sequencing project
Growth Conditions

Temperature: 25°C

Cryopreservation

M9 buffer
KH2PO4, 3.0 g
Na2HPO4, 6.0 g
NaCl, 5.0 g
MgSO4 (1M), 1.0 ml
Distilled H2O, 1.0 L
Dissolve ingredients in 1 L of distilled water. Distribute 200 to 500 ml aliquots into appropriate sized bottles and autoclave for 15 minutes.

Harvest and Preservation

  1. To harvest the Nematocida culture, add 5 ml of M9 buffer to an infected NGM plate and transfer the suspension to a 15 ml centrifuge tube.
  2. Centrifuge at 200 x g for 5 min. Remove the supernatant, resuspend the pellet in 5 ml of M9 buffer, and repeat the centrifugation step.
  3. Repeat step 2 a minimum of five times in order to wash the infected worms.
  4. After the last wash, resupend the pellet in 1 ml of M9 buffer and transfer the suspension to a 2 ml microcentrifuge tube. Add Silicon carbide beads (BioSpec Products, Inc.) to the tube and vortex for 1 minute. Repeat the procedure 4-5 times.  Filter the worm extract through a Whatman filter paper number 1 to remove eggs and any remaining intact worms.
  5. Perform a spore count of the worm extract and adjust the concentration to ≥ 3 x 107 spores/ml.
    NOTE: If the concentration of spores is too low, harvest infected worms from additional NGM plates to yield the desired concentration.
  6. Mix the extract with an equal volume of M9 buffer containing 30% glycerol. The final concentration of the extract will be ≥ 1.5 x 107 spores/ml and 15% glycerol.
  7. Dispense 70 ml aliquots into 1.0-2.0 ml sterile plastic screw-capped cryovials.
  8. Place vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. If freezing unit can compensate for the heat of fusion, maintain rate at -1o C/min through heat of fusion. At -40°C plunge ampules into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus. Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen. (The cooling rate in this apparatus is approximately -1°C/min.)
  9. Store frozen ampules in either the vapor or liquid phase of a nitrogen refrigerator.
Name of Depositor ER Troemel
References

Troemel ER, et al. Microsporidia are natural intracellular parasites of the nematode Caenorhabditis elegans. PLoS Biol. 6: 2736-2752, 2008. PubMed: 19071962

Nematocida parisii ERTm1 genome sequencing project Available from: NCBI BioProject

梅經(jīng)理 17280875617 1438578920
胡經(jīng)理 13345964880 2438244627
周經(jīng)理 17757487661 1296385441
于經(jīng)理 18067160830 2088210172
沈經(jīng)理 19548299266 2662369050
李經(jīng)理 13626845108 972239479
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