Phenotyping protocols

Selection of Protocols for RediMODEL

RediMODEL Protocols - a series of fast and easy-to-use protocols. Selected for clear and quantifiable results. Most assays are designed to be performed in under 24 hrs.

Strain Recovery

Protocol Name

Protocol Link      

Standard Recovery Prep (R001)

more info

Neurological Screens

Protocol Name

Protocol Link      

Low Stimulation Pharynx Pumping (P001)

more info

High Stimulation Pharynx Pumping (P002)

more info

Unstimulated Pharynx Pumping (P003)

more info

Swimming Worm Induced Paralysis (P004)

more info

Hydrogen Peroxide Sensitivity (P007)

more info

Development Screens

Protocol Name

Protocol Link      

Timed Egg Lay (P005)

more info

Brood Size (P006)

more info

List of RediMODEL Kits: RediMODEL Kits

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RediMODEL Protocols

Standard Recovery Protocol (R001)

DESCRIPTION:

The Standard Recovery Protocol has a short incubation period needed to raise RediMODEL animals to adult worm status. Typical length of time to prepare Kit to be ready for phenotyping assay is only 2 days of growth at 22 oC.


Kit Components:

RediMODEL kit contains 3 vials of your desired strains. Typical kits contain the following:

     1. Control: Typical set of controls are:

  • N2 animals for KO kit
  • RediMODEL-KO for KI kit
  • RediMODEL-KO and RediMODEL-KI for CV kit

     2. Test:  Mutant allele

Included in Kit:

  1. Growth plates (6 count)

  2. Wash buffer (30 mL)

  3. Recovery tubes (6 count)

Not included equipment and materials

  1. P1000 pipetor

  2. P1000 pipet tips

  3. Incubator set to 22 C (optional if ambient temperature in near a constant 22 C)

Sample Preparation Protocol:

  1. Remove growth plates and buffer stored at 4 C and place them at room temperature.

  2. Thaw frozen strains at room temperature for ~15 min or until thawed (Caution: rocking back and forth occasionally is ok, but do not vortex or hand thaw).

  3. While strains are thawing, label each of the growth plates on the bottom side, then flip plate back to upright position.

  4. With a P1000 pipet,  pipet thawed strain up and down 3-5 times and transfer 250uL of the liquid to the appropriately labeled plate in the center of the growth media. Swirl plate and spread liquid evenly over the plate. 

  5. Carefully place plates upright in controlled incubation environment that will not be disturbed. Allow them to dry for 4hrs. This may take longer depending on environmental conditions. Allowing them to dry overnight is fine as well. 

  6. Once liquid has dried, flip dishes over and return to controlled incubation environment .

  7. Incubate worms for 2 days.  Caution:  if temperature control was sub-optimal or animals are known to be developmentally delayed, a simple dissecting scope can be useful for assessing appropriate size and age. Many modern cell phones at maximal zoom and manual focus can be adequate for age and size assessment. Strains that display developmental delays in growth are noted. Please see packing sheet for further details.

Recovery:

  1. After worms have reached the desired staging for you assay, add 1 mL of wash buffer with P1000 pipetor to center of each plate.
  2. Swirl gently to dislodge worms from surface.
  3. Suck up worms/buffer with a P1000 and transfer into a labelled recovery tube.
  4. Pellet worms by letting them stand in a rack for 10 min or spinning them in a microcentrifuge for 5 sec at 4k rpm.
  5. Using a P1000, remove most of the buffer, leaving ~50uL of buffer on top of the worm pellet.
  6. Add 950 ul of wash buffer and invert worms 3-5 times.
  7. Repeat pelleting and washing of worms 3 times. 
  8. After the last pelleting of your worms, aspirate all but ~50uL of wash buffer on top of the worms. Make the volumes for all of your samples as close to the same as possible.
  9. Your worms are now ready for your phenotypic assay! 
  10. Go to your specific assay protocol to complete your experiments. 
 

List of RediMODEL Protocols: RediMODEL Protocols

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Low Stimulation Pharynx Pumping Protocol (P001) | $395 per kit Buy Now

RediMODEL Protocols

Low Stimulation Pharynx Pumping Protocol (P001)

DESCRIPTION:

The Low Stimulation Pharynx Pumping Protocol is useful on genetic mutants that exhibit a moderate alteration in pumping rates. Best used when deviation from N2 is 2-fold or less.


Protocol Components:

RediMODEL kit contains 3 vials of your desired strains. Typical kits contain the following:

     1. Control: Typical set of controls are:

  • N2 animals for KO kit
  • RediMODEL-KO for KI kit
  • RediMODEL-KO and RediMODEL-KI for CV kit

     2. Test:  Mutant allele

Included in kit:

  1. Control buffer (2x stock)
  2. Test buffer (2x buffer containing 0.2 mM serotonin)
  3. NemaMetrix chip

Not-included equipment and materials:

  1. ScreenChip System by NemaMetrix

Sample Testing:

  1. After Standard Recovery Protocol is performed for kit prep, make sure that 50 uL of buffer and worms is present in each recovery tube. If volumes vary, add buffer until the levels are the same and roughly 50uL. 
  2. Add 50 uL of Control or Test buffer to each tube of worms and invert 5 times.
  3. Incubate in a tube rack for 30 minutes at room temperature.
  4. Perform ScreenChip as per manufacture protocol.

List of RediMODEL Protocols: RediMODEL Protocols

Discover RediMODEL | Kits starting at $845 ea. RediMODEL kits

Low Stimulation Pharynx Pumping Protocol (P001) | $395 per kit Buy Now

RediMODEL protocol kits: | $395 ea.

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Low Stimulation Pharynx Pumping (P001)

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High Stimulation Pharynx Pumping Protocol (P002) | $395 per kit Buy Now

RediMODEL Protocols

High Stimulation Pharynx Pumping Protocol (P002)

DESCRIPTION:

The High Stimulation Pharynx Pumping Protocol is useful on genetic mutants that exhibit to a significant depression of pharynx pumping rates. Best used when deviation from N2 is more than 2-fold suppressed relative to N2 wild type.


Protocol Components:

RediMODEL kit contains 3 vials of your desired strains. Typical kits contain the following:

     1. Control: Typical set of controls are:

  • N2 animals for KO kit
  • RediMODEL-KO for KI kit
  • RediMODEL-KO and RediMODEL-KI for CV kit

     2. Test:  Mutant allele

Included in kit:

  1. Control buffer (2x stock)
  2. Test buffer (2x buffer containing 0.2 mM serotonin)
  3. NemaMetrix chip

Not-included equipment and materials:

  1. ScreenChip System by NemaMetrix

Sample Testing:

  1. After Standard Recovery Protocol is performed for kit prep, make sure that 50 uL of buffer and worms is present in each recovery tube. If volumes vary, add buffer until the levels are the same and roughly 50uL. 
  2. Add 50 uL of Control or Test buffer to each tube of worms and invert 5 times.
  3. Incubate in a tube rack for 30 minutes at room temperature.
  4. Perform ScreenChip as per manufacture protocol.

List of RediMODEL Protocols: RediMODEL Protocols

Discover RediMODEL | Kits starting at $845 ea. RediMODEL kits

High Stimulation Pharynx Pumping Protocol (P002) | $395 per kit Buy Now

RediMODEL protocol kits: | $395 ea.

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High Stimulation Pharynx Pumping (P002)

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NO Stimulation Pharynx Pumping Protocol (P003) | $395 per kit Buy Now

RediMODEL Protocols

NO Stimulation Pharynx Pumping Protocol (P003)

DESCRIPTION:

The No Stimulation Pharynx Pumping Protocol is useful on genetic mutants that exhibit to a significant overactivation of pharynx pumping rates. Best used when deviation from N2 is more than 2-fold higher pumping rate relative to N2 wild type.


Protocol Components:

RediMODEL kit contains 3 vials of your desired strains. Typical kits contain the following:

     1. Control: Typical set of controls are:

  • N2 animals for KO kit
  • RediMODEL-KO for KI kit
  • RediMODEL-KO and RediMODEL-KI for CV kit

     2. Test:  Mutant allele

Included in kit:

  1. Control buffer (2x stock)
  2. NemaMetrix chip

Not-included equipment and materials:

  1. ScreenChip System by NemaMetrix

Sample Testing:

  1. After Standard Recovery Protocol is performed for kit prep, make sure that 50 uL of buffer and worms is present in each recovery tube. If volumes vary, add buffer until the levels are the same and roughly 50uL. 
  2. Add 50 uL of Control buffer to each tube of worms and invert 5 times.
  3. Incubate in a tube rack for 30 minutes at room temperature.
  4. Perform ScreenChip as per manufacture protocol.

List of RediMODEL Protocols: RediMODEL Protocols

Discover RediMODEL | Kits starting at $845 ea. RediMODEL kits

NO Stimulation Pharynx Pumping Protocol (P003) | $395 per kit Buy Now

RediMODEL protocol kits: | $395 ea.

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Unstimulated Pharynx Pumping (P003)

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Thrashing/Swimming Worm Induced Paralysis (SWIP) Assay Protocol (P004) | $395 per kit Buy Now

RediMODEL Protocols

Thrashing/Swimming Worm Induced Paralysis (SWIP) Assay Protocol (P004)

DESCRIPTION:

The Thrashing/Swimming Worm Induced Paralysis (SWIP) Assay Protocol is useful on genetic mutants that start to exhibit uncoordinated movement after exposure to liquid media. Standard assays assess capacity to thrash normally after 10 minutes in hypotonic solution.


Protocol Components:

RediMODEL kit contains 3 vials of your desired strains. Typical kits contain the following:

     1. Control: Typical set of controls are:

  • N2 animals for KO kit
  • RediMODEL-KO for KI kit
  • RediMODEL-KO and RediMODEL-KI for CV kit

     2. Test:  Mutant allele

Included in kit:

  1. 96-well microplate
  2. Test Buffer (2x stock)

Not-included equipment and materials:

  1. Imaging system (microscope or cell phone camera).

Sample Testing:

  1. After Standard Recovery Protocol is performed for kit prep, confirm 50 uL each is present in tubes for Control and Test worms (If not adjust accordingly).
  2. Add 950 uL of Test Buffer to each tube of worms and invert 5 times.
  3. Invert 5 times and transfer 100ul into each of a 96 well microplate.
  4. Incubate 10 min at room temperature.
  5. Score by visual inspection for ratio of animals showing uncoordinated paralysis phenotype. 

List of RediMODEL Protocols: RediMODEL Protocols

Discover RediMODEL | Kits starting at $845 ea. RediMODEL kits

Swimming Worm Induced Paralysis (SWIP) Protocol (P004) | $395 per kit Buy Now

RediMODEL protocol kits: | $395 ea.

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Swimming Worm Induced Paralysis (P004)

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Timed Egg Lay Protocol (P005) | $395 per kit Buy Now

RediMODEL Protocols

Timed Egg Lay Protocol (P005)

DESCRIPTION:

The Timed Egg Lay Protocol is useful on genetic mutants with defects in egg laying that cause "bag of worms" phenotypes. Standard Assay detects percentage of eggs layed on plate relative to N2 control.


Protocol Components:

RediMODEL kit contains 3 vials of your desired strains. Typical kits contain the following:

     1. Control: Typical set of controls are:

  • N2 animals for KO kit
  • RediMODEL-KO for KI kit
  • RediMODEL-KO and RediMODEL-KI for CV kit

     2. Test:  Mutant allele

Included in kit:

  1. NGM plates without food
  2. Test Buffer

Not-included equipment and materials:

  1. Imaging system (microscope or cell phone camera).

Sample Testing:

  1. After Standard Recovery Protocol is performed for kit prep, confirm 50 ul each is present in tubes for Control and Test worms (If not adjust accordingly).
  2. Add 950 ul of Test Buffer to each tube of worms.
  3. Pipet up and down 5 times and add 50uL of worm solution to a labelled plate.
  4. Incubate 4 hr at room temperature.
  5. Score by visual inspection for number of eggs on plate. 

List of RediMODEL Protocols: RediMODEL Protocols

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Timed Egg Lay Protocol (P005) | $395 per kit Buy Now

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Timed Egg Lay (P005)

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Brood Size Protocol (P006) | $395 per kit Buy Now

RediMODEL Protocols

Brood Size Protocol (P006)

DESCRIPTION:

The Brood Size Protocol (P006) is useful on genetic mutants with defects in numbers of progeny produced. Standard Assay detects percentage of animals on plate relative to N2 control.


Protocol Components:

RediMODEL kit contains 3 vials of your desired strains. Typical kits contain the following:

     1. Control: Typical set of controls are:

  • N2 animals for KO kit
  • RediMODEL-KO for KI kit
  • RediMODEL-KO and RediMODEL-KI for CV kit

     2. Test:  Mutant allele

Included in kit:

  1. NGM plates with food
  2. Test Buffer (30mL)

Not-included equipment and materials:

  1. Imaging system (microscope or cell phone camera)

Sample Testing:

  1. After Standard Recovery Protocol is performed for kit prep, confirm 50 ul each is present in tubes for Control and Test worms (If not adjust accordingly).
  2. Add 950 ul of Test Buffer to each tube of worms.
  3. Pipet up and down 5 times and transfer 25ul onto an NGM plate.
  4. Score and record number of animals transferred.
  5. Incubate 2 days at 22 C.
  6. Score by visual inspection for number of adult animals on plate. 

List of RediMODEL Protocols: RediMODEL Protocols

Discover RediMODEL | Kits starting at $845 ea. RediMODEL kits

Brood Size Protocol (P006) | $395 per kit Buy Now

RediMODEL protocol kits: | $395 ea.

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Brood Size (P006)

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Hydrogen Peroxide Sensitivity Protocol (P007) | $395 per kit Buy Now

RediMODEL Protocols

Hydrogen Peroxide Sensitivity Protocol (P007)

DESCRIPTION:

The Hydrogen Peroxide Sensitivity Protocol is useful on genetic mutants that exhibit a sensitivity to oxidative stress conditions.


Protocol Components:

RediMODEL kit contains 3 vials of your desired strains. Typical kits contain the following:

     1. Control: Typical set of controls are:

  • N2 animals for KO kit
  • RediMODEL-KO for KI kit
  • RediMODEL-KO and RediMODEL-KI for CV kit

     2. Test:  Mutant allele

Included in kit:

  1. Test Buffer (30 ml)
  2. 15 ml conical tube
  3. 96-well microplate
  4. NemaMetrix ScreenChip

Not-included equipment and materials:

  1. ScreenChip System by NemaMetrix
  2. Hydrogen Peroxide (30%)

Sample Testing:

  1. After Standard Recovery Protocol is performed for kit prep, confirm 50 uL each is present in tubes for Control and Test worms (If not adjust accordingly).
  2. Add 100 uL of Test Buffer to worms to make 150 uL final.
  3. To the 15 mL conical, add 10 mL of Test Buffer.
  4. Add 20.5 uL of 30% hydrogen peroxide to the 15 mL conical.
  5. Cap and mix solution in 15 mL conical.
  6. Transfer 50 uL of solution in 15 ml conical to appropriate position in microwell plate (every other well).
  7. Add  50 uL of Test Buffer to intervening wells as control.
  8. Transfer 50 ul of worms to appropriate position of microwell plate (each tube of worms is used twice - pipet up down 5 x before recovering solution to suspend worms).
  9. incubate 120 min at rt.
  10. Perform ScreenChip as per manufacture protocol  (suggested each tube set is done at 90 animals each for 2 min measurement interval each).

List of RediMODEL Protocols: RediMODEL Protocols

Discover RediMODEL | Kits starting at $845 ea. RediMODEL kits

Hydrogen Peroxide Sensitivity Protocol (P007) | $395 per kit Buy Now

RediMODEL protocol kits: | $395 ea.

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Hydrogen Peroxide Sensitivity (P007)

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RediMODEL protocol kits: | $395 ea.

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New protocol

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