Difference between revisions of "Team:TUDelft/Wetlab/Protocols"

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               <button class="collapsible cadpbl">List of laboratory guidelines</button>
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               <button class="collapsible cadpbl">Colony PCR</button>
 
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                    <li>At any given time, individuals should wear minimal protective clothes (closed shoes, long sleeve shirts, long trouser legs, white lab coat). Depending on the experimental conditions, additional measures like gloves or protective eyewear might be needed;</li>
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                    <li>When entering and leaving the BSL-1 laboratory space, individuals should wash their hands to prevent any unwanted spreading of contained organisms.</li>
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<li>Under sterile conditions, pick a colony and dilute it in 10 µL of milli-Q water. <br>
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                    <li>The working space is kept organized, tidy and clean;</li>
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NOTE: After you pick the colony, it cannot be used again. It is therefore recommended to make a 'back-up'-plate where you grow the colonies again. This plate should be incubated overnight at 37 °C. </li>
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                    <li>Eating, drinking, smoking, presence/storage of nutritious material for consumption, application of cosmetics or contact lenses is prohibited.</li>
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<li>Incubate at 90 °C for 10 min. <br>
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                    <li>Pipetting with your mouth is prohibited;</li>
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NOTE: Instead of separate 'cooking' of the cells before the PCR, this step can be incorporated in the PCR program. The initial denaturation step at 98 °C should then be prolonged to 5 minutes (300 seconds). </li>
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                    <li>Presence of vermin is strictly prohibited;</li>
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<li>Prepare the GoTaq master mix for all the samples in a single 1.5 mL tube (NOTE: mind pipetting errors so prepare at little bit more master mix!). For one sample: </li>
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                    <li>Jackets, coats, bags, sweaters etc. (and other likewise personal belongings) should be stored outside the working space;</li>
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                    <li>In case of contamination of surfaces, these should be cleaned and desinfected instantly;</li>
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                    <li>Contaminated clothing should be directly autoclaved in case of contamination with biological agents/ GMOs through spilling.</li>
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                    <li>Contaminated waste should be directed to designated areas for appropriate treatment thereof.</li>
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Revision as of 07:59, 15 September 2018

Wetlab Protocols

Text to write to introduce the protocols


  1. Under sterile conditions, pick a colony and dilute it in 10 µL of milli-Q water.
    NOTE: After you pick the colony, it cannot be used again. It is therefore recommended to make a 'back-up'-plate where you grow the colonies again. This plate should be incubated overnight at 37 °C.
  2. Incubate at 90 °C for 10 min.
    NOTE: Instead of separate 'cooking' of the cells before the PCR, this step can be incorporated in the PCR program. The initial denaturation step at 98 °C should then be prolonged to 5 minutes (300 seconds).
  3. Prepare the GoTaq master mix for all the samples in a single 1.5 mL tube (NOTE: mind pipetting errors so prepare at little bit more master mix!). For one sample: