Difference between revisions of "Team:Oxford/Test/Beads"

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                             </p>
 
                             </p>
 
                       </div>
 
                       </div>
−
                            <div id="11-08-2015-Method">
+
                        <div id="11-08-2015-Method">
−
                                <h3>Method</h3>
+
                            <h3>Method</h3>
−
                                    <p>In a fume cupboard, break up a standard (polystyrene) petri dish into small pieces and dissolve in the minimum amount of ethyl acetate.</p>
+
                                <p>In a fume cupboard, break up a standard (polystyrene) petri dish into small pieces and dissolve in the minimum amount of ethyl acetate.</p>
−
                                    <p>Prepare a 150 mL 1.5 % Agarose solution in breaker with screw cap. Microwave on high for 2 minutes and then cool to 40 °C in water bath, when microwaving ensure the screw cap is placed on loosely.</p>
+
                                <p>Prepare a 150 mL 1.5 % Agarose solution in breaker with screw cap. Microwave on high for 2 minutes and then cool to 40 °C in water bath, when microwaving ensure the screw cap is placed on loosely.</p>
−
                                    <p>Remove agarose solution from water bath and bring to laminar airflow cupboard. Pour the agarose into petri dish to a depth of 1cm. Allow to set, this should take roughly 15 minutes.</p>
+
                                <p>Remove agarose solution from water bath and bring to laminar airflow cupboard. Pour the agarose into petri dish to a depth of 1cm. Allow to set, this should take roughly 15 minutes.</p>
−
                                    <p>Still under laminar flow, use an autoclaved 1 cm diameter hole borer to core out the required number of identical agarose cylinders and place in a second petri dish and allow drying.</p>
+
                                <p>Still under laminar flow, use an autoclaved 1 cm diameter hole borer to core out the required number of identical agarose cylinders and place in a second petri dish and allow drying.</p>
−
                                    <p>Bring uncoated ‘beads’ to fume cupboard. Using autoclaved needles, pick up individual beads and dip in ethyl acetate-polystyrene solution, stand each bead on needle upright in fume cupboard on blob of blu-tac to allow ethyl acetate evaporation and the coating to set.  When the coating is almost set the beads should be able to be handled through gloves without damaging the coat. Remove needle and mould the polymer coat over the needle hole, sealing them.</p>
+
                                <p>Bring uncoated ‘beads’ to fume cupboard. Using autoclaved needles, pick up individual beads and dip in ethyl acetate-polystyrene solution, stand each bead on needle upright in fume cupboard on blob of blu-tac to allow ethyl acetate evaporation and the coating to set.  When the coating is almost set the beads should be able to be handled through gloves without damaging the coat. Remove needle and mould the polymer coat over the needle hole, sealing them.</p>
−
                            </div>
+
                        </div>
−
                            <div id="11-08-2015-COSHH">
+
                        <div id="11-08-2015-COSHH">
−
                                <h3>Control of substances Hazardous to Health (COSHH) Assessment</h3>
+
                            <h3>Control of substances Hazardous to Health (COSHH) Assessment</h3>
−
                                    <h4>Agarose</h4>
+
                                <h4>Agarose</h4>
−
                                    <ul>
+
                                <ul>
−
                                        <li>Not a hazardous substance</li>
+
                                    <li>Not a hazardous substance</li>
−
                                    </ul>
+
                                </ul>
−
                                    <h4>Ethyl Acetate</h4>
+
                                <h4>Ethyl Acetate</h4>
−
                                    <ul>
+
                                <ul>
−
                                        <li>H225 – Highly flammable liquid and vapour.</li>
+
                                    <li>H225 – Highly flammable liquid and vapour.</li>
−
                                        <li>H319 – Causes serious eye irritation.</li>
+
                                    <li>H319 – Causes serious eye irritation.</li>
−
                                        <li>H336 – May cause drowsiness or dizziness.></li>
+
                                    <li>H336 – May cause drowsiness or dizziness.></li>
−
                                        <li>P210 – Keep away from heat, hot surfaces, sparks, open flames and other ignition sources. No smoking.</li>
+
                                    <li>P210 – Keep away from heat, hot surfaces, sparks, open flames and other ignition sources. No smoking.</li>
−
                                        <li>P261 – Avoid breathing vapours.</li>
+
                                    <li>P261 – Avoid breathing vapours.</li>
−
                                        <li>P305+P351+P338 – IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do so. Continue rinsing.</li>
+
                                    <li>P305+P351+P338 – IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do so. Continue rinsing.</li>
−
                                    </ul>
+
                                </ul>
−
                                    <p>Ethyl acetate poses the biggest safety hazard in this experiment so therefore do all steps involving it in the fumehood. Be careful of cutting self when breaking up the Petri dish, as the pieces can be sharp.</p>
+
                                <p>Ethyl acetate poses the biggest safety hazard in this experiment so therefore do all steps involving it in the fumehood. Be careful of cutting self when breaking up the Petri dish, as the pieces can be sharp.</p>
−
                                </div>
+
                            </div>
−
                                <div id="11-08-2015-Write-Up">
+
                            <div id="11-08-2015-Write-Up">
−
                                    <h3>Write Up</h3>
+
                                <h3>Write Up</h3>
−
                                    <p>150mL of 1.5% agarose was made up using 2.25g of agarose powder and 150 mL of MilliQ.</p>
+
                                <p>150mL of 1.5% agarose was made up using 2.25g of agarose powder and 150 mL of MilliQ.</p>
−
                                    <p>In a fumehood, one standard petri dish was broken into small pieces and placed into a 500mL beaker. To this, 50 mL of ethyl acetate was added along with a stirrer bar and the mixture was left to stir for 10 minutes. After 10 minutes the petri dish was still not dissolved so a further 10 mL of ethyl acetate was added and left to stir again.</p>
+
                                <p>In a fumehood, one standard petri dish was broken into small pieces and placed into a 500mL beaker. To this, 50 mL of ethyl acetate was added along with a stirrer bar and the mixture was left to stir for 10 minutes. After 10 minutes the petri dish was still not dissolved so a further 10 mL of ethyl acetate was added and left to stir again.</p>
−
                                    <p>The agarose was then poured into a petri dish to a depth of 1cm and left to set for 15 minutes. After this was set, using an eppendorf tube with the lib cut off cylinders of agarose were cut out and placed into a second petri dish.</p>
+
                                <p>The agarose was then poured into a petri dish to a depth of 1cm and left to set for 15 minutes. After this was set, using an eppendorf tube with the lib cut off cylinders of agarose were cut out and placed into a second petri dish.</p>
−
                                    <p>After the agarose was left to dry it was taken to the fumehood, to be coated. The agarose beads were placed onto the end of pipette tips and then dipped into the ethyl acetate-polystyrene mixture. These were then left to dry standing up on the pipette tips.</p>
+
                                <p>After the agarose was left to dry it was taken to the fumehood, to be coated. The agarose beads were placed onto the end of pipette tips and then dipped into the ethyl acetate-polystyrene mixture. These were then left to dry standing up on the pipette tips.</p>
−
                                    <p>The beads were left to dry overnight and it was found that the coating did not stick sufficiently to the agarose.</p>
+
                                <p>The beads were left to dry overnight and it was found that the coating did not stick sufficiently to the agarose.</p>
−
                                <div id="11-08-2015-Findings">
+
                            <div id="11-08-2015-Findings">
−
                                    <h3>Findings</h3>
+
                                <h3>Findings</h3>
−
                                    <ul>
+
                                <div class="image image-right">
−
                                        <li>Beads are far too big to fit into a catheter, but could be used in larger pipes.</li>
+
                                    <img src="https://static.igem.org/mediawiki/2015/d/d8/Ox_Agarosebeads.jpg">
−
                                        <li> It is very hard to get an even coating of the ethyl acetate-polystyrene mixture around the bead and it would tend to slide off.</li>
+
                                    <p>Finished Beads</p>
−
                                        <li>Ethyl acetate-Polystyrene mixture was difficult to clean up.</li>
+
−
                                        <li>Is the coating actually porous?</li>
+
−
                                    </ul>
+
−
                                    <div class="image image-right">
+
−
                                        <img src="https://static.igem.org/mediawiki/2015/d/d8/Ox_Agarosebeads.jpg">
+
−
                                        <p>Finished Beads</p>
+
−
                                    </div>
+
 
                                 </div>
 
                                 </div>
 +
                                <ul>
 +
                                    <li>Beads are far too big to fit into a catheter, but could be used in larger pipes.</li>
 +
                                    <li> It is very hard to get an even coating of the ethyl acetate-polystyrene mixture around the bead and it would tend to slide off.</li>
 +
                                    <li>Ethyl acetate-Polystyrene mixture was difficult to clean up.</li>
 +
                                    <li>Is the coating actually porous?</li>
 +
                                </ul>
 
                             </div>
 
                             </div>
 
                         </div>
 
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{{:Team:Oxford/Templates/Foot}}
 
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Revision as of 10:05, 13 August 2015

Beads Notebook

11/08/2015

Aim

To improve on last years Oxford iGEM team biobead design, using agarose to make up beads and then coating them with a ethyl acetate and polystyrene mixture.

Method

In a fume cupboard, break up a standard (polystyrene) petri dish into small pieces and dissolve in the minimum amount of ethyl acetate.

Prepare a 150 mL 1.5 % Agarose solution in breaker with screw cap. Microwave on high for 2 minutes and then cool to 40 °C in water bath, when microwaving ensure the screw cap is placed on loosely.

Remove agarose solution from water bath and bring to laminar airflow cupboard. Pour the agarose into petri dish to a depth of 1cm. Allow to set, this should take roughly 15 minutes.

Still under laminar flow, use an autoclaved 1 cm diameter hole borer to core out the required number of identical agarose cylinders and place in a second petri dish and allow drying.

Bring uncoated ‘beads’ to fume cupboard. Using autoclaved needles, pick up individual beads and dip in ethyl acetate-polystyrene solution, stand each bead on needle upright in fume cupboard on blob of blu-tac to allow ethyl acetate evaporation and the coating to set. When the coating is almost set the beads should be able to be handled through gloves without damaging the coat. Remove needle and mould the polymer coat over the needle hole, sealing them.

Control of substances Hazardous to Health (COSHH) Assessment

Agarose

  • Not a hazardous substance

Ethyl Acetate

  • H225 – Highly flammable liquid and vapour.
  • H319 – Causes serious eye irritation.
  • H336 – May cause drowsiness or dizziness.>
  • P210 – Keep away from heat, hot surfaces, sparks, open flames and other ignition sources. No smoking.
  • P261 – Avoid breathing vapours.
  • P305+P351+P338 – IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do so. Continue rinsing.

Ethyl acetate poses the biggest safety hazard in this experiment so therefore do all steps involving it in the fumehood. Be careful of cutting self when breaking up the Petri dish, as the pieces can be sharp.

Write Up

150mL of 1.5% agarose was made up using 2.25g of agarose powder and 150 mL of MilliQ.

In a fumehood, one standard petri dish was broken into small pieces and placed into a 500mL beaker. To this, 50 mL of ethyl acetate was added along with a stirrer bar and the mixture was left to stir for 10 minutes. After 10 minutes the petri dish was still not dissolved so a further 10 mL of ethyl acetate was added and left to stir again.

The agarose was then poured into a petri dish to a depth of 1cm and left to set for 15 minutes. After this was set, using an eppendorf tube with the lib cut off cylinders of agarose were cut out and placed into a second petri dish.

After the agarose was left to dry it was taken to the fumehood, to be coated. The agarose beads were placed onto the end of pipette tips and then dipped into the ethyl acetate-polystyrene mixture. These were then left to dry standing up on the pipette tips.

The beads were left to dry overnight and it was found that the coating did not stick sufficiently to the agarose.

Findings

Finished Beads

  • Beads are far too big to fit into a catheter, but could be used in larger pipes.
  • It is very hard to get an even coating of the ethyl acetate-polystyrene mixture around the bead and it would tend to slide off.
  • Ethyl acetate-Polystyrene mixture was difficult to clean up.
  • Is the coating actually porous?