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<div class = "protocols" id="protocol"> | <div class = "protocols" id="protocol"> | ||
<div class = "prot" id="prot1"> Making LB Plates </div> | <div class = "prot" id="prot1"> Making LB Plates </div> | ||
− | <div class = "num" id="1"> This is the protocol for making LB gels. The measurements given can make approximately 24 plates. If fewer plates are wanted, the measurements can be halved to make approximately 10-12 gels.<br>Materials <ul><li>Large bottle with a screw-on lid </li> | + | <div class = "num" id="1"> This is the protocol for making LB gels. The measurements given can make approximately 24 plates. If fewer plates are wanted, the measurements can be halved to make approximately 10-12 gels.<br><p style="font-weight:bold">Materials </p><ul><li>Large bottle with a screw-on lid </li> |
<li>Stir bar & Stir plate </li> | <li>Stir bar & Stir plate </li> | ||
<li>DI H2O </li> | <li>DI H2O </li> | ||
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</ul> | </ul> | ||
<br> | <br> | ||
− | Procedure<br> | + | <p style="font-weight:bold"> |
+ | Procedure</p><br> | ||
Fill a large bottle with lid with 570 mL of DI H2O. Insert a stir bar and place the bottle on the stir plate. Mix in 6g Tryptone, 3g yeast extract, 9g agar, and 6g NaCl. Ensure the solution has been mixed thoroughly and standardize the pH meter with the neutral standard solution. Add 2M NaOH to the solution dropwise until the pH reaches 7. <br> <br> Check the water level in the autoclave and put autoclave tape on the bottle. Ensure the lid is on but not tightened. Put the bottle in the autoclave and tighten the door. Set it for the liquid cycle and wait until the cycle is over and the pressure reaches 0 to open the door.<br><br> Put the bottle back on the stir plate and turn it to a low setting to prevent bubbles forming. Let the mixture cool until you can touch the bottle for several seconds comfortably. Add 600 µL of the chosen antibiotic (at 50mg/mL) to the bottle. Label all plates before you begin to pour. Wearing a glove, use the aseptic technique to pour the media into the plates and leave them to solidify. | Fill a large bottle with lid with 570 mL of DI H2O. Insert a stir bar and place the bottle on the stir plate. Mix in 6g Tryptone, 3g yeast extract, 9g agar, and 6g NaCl. Ensure the solution has been mixed thoroughly and standardize the pH meter with the neutral standard solution. Add 2M NaOH to the solution dropwise until the pH reaches 7. <br> <br> Check the water level in the autoclave and put autoclave tape on the bottle. Ensure the lid is on but not tightened. Put the bottle in the autoclave and tighten the door. Set it for the liquid cycle and wait until the cycle is over and the pressure reaches 0 to open the door.<br><br> Put the bottle back on the stir plate and turn it to a low setting to prevent bubbles forming. Let the mixture cool until you can touch the bottle for several seconds comfortably. Add 600 µL of the chosen antibiotic (at 50mg/mL) to the bottle. Label all plates before you begin to pour. Wearing a glove, use the aseptic technique to pour the media into the plates and leave them to solidify. | ||
Revision as of 16:39, 18 July 2018