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| | <a href="#">HOME > RESEARCH > SAFETY > PROTOCOLS</a> | | <a href="#">HOME > RESEARCH > SAFETY > PROTOCOLS</a> |
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| − | <h3>Steaking/Plating Microbial Cultures on Agar</h3> | + | <h3><a href="#">Inoculating Cultures in Liquid Media</a></h3> |
| − | <p><b>Goal:</b> To isolate and grow isolated cultures of <i>E. coli</i> on Agar plates<p>
| + | <h3><a href="#">Lysogeny Broth Preparation</a></h3> |
| − | <p><b>Materials:</b><br>Bunsen burner<br>Disposable loops used to streak plates Lysogeny broth(LB) agar plates (with antibiotics if required)<br>Culture of E. coli (in plate or in shake flask/test tube)</p> | + | <h3><a href="#">Lysogeny Agar Plate Preparation</a></h3> |
| − | <p><b>Procedure:</b><br>1. Ignite the Bunsen burner to maintain a sterile environment<br>2. If culture is in a petri dish: Use the tip of a disposable loop and pick up an isolated culture<br>3. If culture is in a shake flask/test tube: Dip the circular end of the loop into the liquid medium culture<br>4. Close the petri dish or shake flask/test tube<br>5. Transfer the collected sample to the LB agar plates by gently touching the tip/loop end with bacteria on the agar and brush over the plate. Make sure to streak at an angle<br>6. Two streaking methods are possible: Brushing the loop all over the plate once without crossing previous streaks, or brushing the loop over three different thirds of the plate<br>7. Allow streaks to dry before labelling each plate and incubating them at the appropriate temperature</p> | + | <h3><a href="#">Chemical Transformations</a></h3> |
| − | <h3>Inoculating Cultures in Liquid Media</h3>
| + | <h3><a href="#">PCR</a></h3> |
| − | <p><b>Goal:</b> To isolate and grow individual colonies of <i>E. coli</i> in a nutrient-rich media<p> | + | <h3><a href="#">Restriction Digestion</a></h3> |
| − | <p><b>Materials:</b><br>Test tubes/Shake flask<br>Media (Lysogeny Broth)<br>Petri dishes containing <i>E. coli</i> culture<br>Pipettes<br>Antibiotics (if required)<br>Bunsen burner</p> | + | <h3><a href="#">Ligation of PCR Product into Plasmid</a></h3> |
| − | <p><b>Procedure:</b><br>1. Ignite the Bunsen burner to maintain a sterile environment<br>2. Obtain a shake flask/test tube and transfer the desired amount of media into the flask/tube. If required, add the appropriate amount of antibiotics<br>3. Open the petri dishes containing the E. coli cultures. Use a pipette tip and carefully pick a single colony from the plate<br>4. Transfer the colony on the pipette tip to the media in the flask/tube<br>5. Label the flask/tube and incubate in a shaker at an appropriate temperature</p> | + | <h3><a href="#">DNA Electrophoresis</a></h3> |
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