Difference between revisions of "Team:Stony Brook/Experiments"

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<h3> NEB Monarch Nucleic Acid Purification Protocols </h3>
 
<h3> NEB Monarch Nucleic Acid Purification Protocols </h3>
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<p> The following NEB kits were used and protocols can be found on their <a href="https://www.neb.com/monarch/monarch-nucleic-acid-purification-kits"> website </a></p>
 
<p> The following NEB kits were used and protocols can be found on their <a href="https://www.neb.com/monarch/monarch-nucleic-acid-purification-kits"> website </a></p>
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<li> Monarch Plasmid Miniprep Kit </li>
 
<li> Monarch Plasmid Miniprep Kit </li>

Revision as of 22:01, 21 July 2016

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Protocols & Experiments

Universal Lab Protocols

Agarose Gel Preparation

  • Add 50mL of 1X TAE buffer to a 500mL erlenmeyer flask
  • Add 0.5g of 1% agarose
  • Microwave for three 20 second intervals and cool slightly
  • Add 1ul Ethidium Bromide
  • Mix and pour into gel box. Add Comb
  • Cool for 30 minutes

YPD Media

  • For liquid media
    • Add 20g of Bacto peptone, 10g of yeast extract and 950mL of water to a flask
    • Autoclave on the liquid setting
    • Add 50mL of 40% glucose and mix
  • For solid media
    • Add same reagents as liquid, plus 24g of Bacto agar
    • Autoclave
    • Stir on a magnetic stir plate and add 50ml of 40% glucose
    • Pour into petri dishes

LB Broth

Add items to flask:

  • 5g Tryptone
  • 2.5g Yeast Extract
  • 5g NaCl
  • H2O filled to 500mL
  • Mix with magnetic stir bar on low heat, autoclave for liquids



Nanodrop

  • Select Nucleic Acid
  • Clean, drop 1ul water, measure
  • Blank
  • Measure with 1ul of substance
  • For PCRs: 260/280 ratio should be near 2

LB Agar

Add items to flask:

  • 5g Tryptone
  • 2.5g Yeast Extract
  • 5g NaCl
  • 7.5g Agar
  • H2O filled to 500mL
  • Mix with magnetic stir bar on low heat, autoclave for liquids


    • NEB Monarch Nucleic Acid Purification Protocols

    The following NEB kits were used and protocols can be found on their website

    • Monarch Plasmid Miniprep Kit
    • Monarch DNA Gel Extraction Kit
    • Monarch PCR & DNA Cleanup Kit
Inspiration