Team:Oxford/Chemicals

iGEM Oxford 2016 - Cure for Copper

Chemicals

A variety of different chemicals were used for the execution of the wet lab experiments. The following list contains all the major chemicals used in our copper chelation system and delivery mechanism studies. Hazardous or dangerous chemicals have also a list of precautionary and hazardous statements as defined by the Globally Harmonized System of Classification and Labelling of Chemicals (GHS)

Ethidium Bromide

Use: Staining DNA electrophoresis gels.

Signal word: Danger


Hazard statement(s)

  • H302 Harmful if swallowed.
  • H330 Fatal if inhaled.
  • H341 Suspected of causing genetic defects.

Precautionary statement(s)

  • P260 Do not breathe dust/fume/gas/mist/vapours/spray.
  • P281 Use personal protective equipment as required.
  • P284 Wear respiratory protection.
  • P310 Immediately call a POISON CENTER or doctor/ physician.

TBE Buffer pH 8.3

Use: General buffer for DNA experiments.
Made of three chemicals: Tris, borate and EDTA

Non-Hazardous


Copper Sulphate Pentahydrate

Use: Induction of copper promoters and Cu2+ chelation assays.

Signal word: Warning


Hazard statement(s)

  • H302 Harmful if swallowed.
  • H315 Causes skin irritation.
  • H319 Causes serious eye irritation.
  • H410 Very toxic to aquatic life with long lasting effects.

Precautionary statement(s)

  • P273 Avoid release to the environment.
  • P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes.Remove contact lenses, if present and easy to do. Continue rinsing.
  • P501 Dispose of contents/container to an approved waste disposal plant.

Agarose

Use: DNA electrophoresis gel matrix

Non-Hazardous


L(+)-Arabinose

Use: Induction of pBAD expression

Non-Hazardous


Bathocuproinedisulfonic Acid Disodium Salt

Use: Chromophore in Cu+/Cu2+ detection assay

Non-Hazardous


L(+)-Sodium Ascorbate

Use: Reducing agent in Cu+/Cu2+ detection assay

Non-Hazardous


L-Glutathione

Use: Reducing agent in Cu+/Cu2+ detection assay.

Non-Hazardous


Chloramphenicol

Use: Antibiotic and positive selection marker for all plasmids except pBAD

Signal word: Danger


Hazard statement(s)

  • H350 May cause cancer

Precautionary statement(s)

  • P201 Obtain special instructions before use.
  • P308 + P313 If exposed or concerned: Get medical advice/attention.

Ampicillin

Use: Antibiotic and positive selection marker for pBAD.

Signal word: Danger


Hazard statement(s)

  • H315 Causes skin irritation
  • H317 May cause an allergic skin reaction
  • H319 Causes serious eye irritation
  • H334 May cause allergy or asthma symptoms or breathing difficulties if inhaled
  • H335 May cause respiratory irritation

Precautionary statement(s)

  • P261 Avoid breathing dust/fume/gas/mist/vapours/spray
  • P280 Wear protective gloves
  • P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing
  • P342 + P311 If experiencing respiratory symptoms: Call a POISON CENTER or doctor/physician

Tris pH 7.0 and 8.0

Use: General buffer for protein and copper assays

Non-Hazardous


Glycerol

Use: Component of protein buffer for protein stability at low temperatures. Used in protein purification lysis and elution buffers. Also, used in SDS-PAGE run buffer

Non-Hazardous


DL-Dithiothreitol

Use: Reducing agent in Cu+/Cu2+ detection assay and protein purification

Signal word: Warning


Hazard statement(s)

  • H302 Harmful if swallowed
  • H315 Causes skin irritation
  • H319 Causes serious eye irritation
  • H335 May cause respiratory irritation

Precautionary statement(s)

  • P261 Avoid breathing dust
  • P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing

Imidazole

Use: Elution of His-tagged target proteins from nickel bead column during protein purification

Signal word: Danger


Hazard statement(s)

  • H302 Harmful if swallowed
  • H314 Causes severe skin burns and eye damage
  • H360D May damage the unborn child

Precautionary statement(s)

  • P201 Obtain special instructions before use
  • P280 Wear protective gloves/protective clothing/eye protection/face protection
  • P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing
  • P310 Immediately call a POISON CENTER or doctor/physician

Nickel Affinity Beads

Use: Selection of His-tagged target proteins during protein purification

Non-Hazardous



Sodium Alginate

Use: Major component of bacterial delivery beads

Non-Hazardous


Crystal Violet

Use: Marker for bead quality assay

Signal word: Danger


Hazard statement(s)

  • H302 Harmful if swallowed
  • H318 Causes serious eye damage
  • H350 May cause cancer
  • H410 Very toxic to aquatic life with long lasting effects

Precautionary statement(s)

  • P201 Obtain special instructions before use
  • P273 Avoid release to the environment
  • P280 Wear protective gloves/ eye protection/ face protection
  • P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing
  • P308 + P313 If exposed or concerned: Get medical advice/attention
  • P501 Dispose of contents/container to an approved waste disposal plant

Lysogeny Broth

Use: Medium for general bacterial growth

Non-Hazardous


Phosphate Buffered Saline pH 7.4

Use: Dilution buffer for flow cytometry experiments

Non-Hazardous


Sodium Dodecyl Sulphate

Use: Detergent in protein identification gels

Signal word: Danger


Hazard statement(s)

  • H228 Flammable solid
  • H302 + H332 Harmful if swallowed or if inhaled
  • H315 Causes skin irritation
  • H318 Causes serious eye damage
  • H335 May cause respiratory irritation
  • H412 Harmful to aquatic life with long lasting effects

Precautionary statement(s)

  • P210 Keep away from heat, hot surfaces, sparks, open flames and other ignition sources. No smoking
  • P261 Avoid breathing dust/fume/gas/mist/vapours/spray
  • P280 Wear eye protection/face protection
  • P301 + P312 + P330 IF SWALLOWED: Call a POISON CENTER/doctor if you feel unwell. Rinse mouth
  • P305 + P351 + P338 + P310 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. Immediately call a POISON CENTER/doctor
  • P370 + P378 In case of fire: Use dry powder or dry sand to extinguish

Ethanol 100%

Use: solvent for chloramphenicol

Signal word: Danger


Hazard statement(s)

  • H225 Highly flammable liquid and vapour
  • H319 Causes serious eye irritation

Precautionary statement(s)

  • P210 Keep away from heat, hot surfaces, sparks, open flames and other ignition sources. No smoking
  • P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing
  • P370 + P378 In case of fire: Use dry powder or dry sand to extinguish
  • P403 + P235 Store in a well-ventilated place. Keep cool

Transfection Buffer I (Manganese[II] Chloride)

Use: First soaking buffer in creating competent E. coli cells

Signal word: Danger


Hazard statement(s)

  • H302 Harmful if swallowed
  • H318 Causes serious eye damage
  • H373 May cause damage to organs (Brain) through prolonged or repeated exposure if inhaled
  • H411 Toxic to aquatic life with long lasting effects

Precautionary statement(s)

  • P273 Avoid release to the environment
  • P280 Wear eye protection/ face protection
  • P301 + P312 + P330 IF SWALLOWED: Call a POISON CENTER or doctor/physician if you feel unwell. Rinse mouth
  • P305 + P351 + P338 + P310 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. Immediately call a POISON CENTER or doctor/physician
  • P391 Collect spillage
  • P501 Dispose of contents/container to an approved waste disposal plant

Transfection Buffer II

Use: Second soaking buffer in creating competent E. coli cells

Non-Hazardous

Chitosan

Use: Coating for bacterial delivery beads

Non-Hazardous


InstantBlue

Use: Protein stain for protein identification gels

Signal word: Warning


Hazard statement(s)

  • H290 May be corrosive to metals
  • H315 Causes skin irritation
  • H319 Causes serious eye irritation

Precautionary statement(s)

  • P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing

NEB Purple Loading Dye

Use: Stabilizer and marker for DNA gel electrophoresis

Non-Hazardous


Roche cOmplete EDTA-free Protease Inhibitor Cocktail

Use: Prevention of protein cleavage during protein purification

Signal word: Danger


Hazard statement(s)

  • H314 Causes severe skin burns and eye damage Skin corrosion/irritation

Precautionary statement(s)

  • P260 Do not breathe dust/fume/gas/mist/vapours/spray
  • P280 Wear protective gloves/protective clothing/eye protection/face protection
  • P301 + P330 + P331 IF SWALLOWED: Rinse mouth. Do NOT induce vomiting
  • P303 + P361 + P353 IF ON SKIN (or hair): Remove/Take off Immediately all contaminated clothing. Rinse SKIN with water/shower
  • P304 + P340 + P310 IF INHALED: Remove victim to fresh air and Keep at rest in a position comfortable for breathing. Immediately call a POISON CENTER or doctor/physician
  • P305 + P351 + P338 + P310 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. Immediately call a POISON CENTER or doctor/physician

Hydrochloric Acid

Use: Simulation of stomach acid conditions in bead experiments

Signal word: Warning


Hazard statement(s)

  • H290 May be corrosive to metals

Sodium Hydroxide

Use: Simulation of duodenum conditions in bead experiments

Signal word: Danger


Hazard statement(s)

  • H290 May be corrosive to metals
  • H314 Causes severe skin burns and eye damage

Precautionary statement(s)

  • P260 Do not breathe dust/fume/gas/mist/vapours/spray
  • P280 Wear protective gloves/protective clothing/eye protection/face protection
  • P303 + P361 + P353 IF ON SKIN (or hair): Take off immediately all contaminated clothing. Rinse skin with water/shower
  • P304 + P340 + P310 IF INHALED: Remove person to fresh air and keep comfortable for breathing. Immediately call a POISON CENTER/doctor
  • P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing

Monobasic Sodium Phosphate

Use: Simulation of duodenum conditions in bead experiments

Non-Hazardous


Glacial Acetic Acid

Use: Bead experiments

Signal word: Danger


Hazard statement(s)

  • H226 Flammable liquid and vapour
  • H314 Causes severe skin burns and eye damage

Precautionary statement(s)

  • P210 Keep away from heat, hot surfaces, sparks, open flames and other ignition sources. No smoking
  • P260 Do not breathe dust/fume/gas/mist/vapours/spray
  • P280 Wear protective gloves/protective clothing/eye protection/face protection
  • P303 + P361 + P353 IF ON SKIN (or hair): Take off immediately all contaminated clothing. Rinse skin with water/shower
  • P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing
  • P370 + P378 In case of fire: Use dry powder or dry sand to extinguish

Calcium Chloride

Use: Forming beads from alginate. Ca2+ causes alginate phase change

Signal word: Warning


Hazard statement(s)

  • H319 Causes serious eye irritation

Precautionary statement(s)

  • P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing