UW-Stout/UV Light SP22 - Revision history

Teagueb: /* Knock-out Protocol */

2022-05-03T19:55:10Z

‎Knock-out Protocol

Teagueb: /* Knock-out Protocol */

2022-05-03T19:54:04Z

‎Knock-out Protocol

Teagueb: /* Knock-out Protocol */

2022-05-03T19:53:35Z

‎Knock-out Protocol

Teagueb: /* Calibration Protocol */

2022-05-03T19:53:01Z

‎Calibration Protocol

Teagueb: /* Knock-out Protocol */

2022-05-03T19:52:54Z

‎Knock-out Protocol

Teagueb: /* Calibration Experiment 2 */

2022-05-03T19:51:54Z

‎Calibration Experiment 2

Teagueb: /* Calibration Experiment 1 */

2022-05-03T19:51:30Z

‎Calibration Experiment 1

Teagueb: /* Equipment */

2022-05-03T19:47:05Z

‎Equipment

Teagueb: /* Calibration Experiment 1 Results */

2022-05-03T19:46:05Z

‎Calibration Experiment 1 Results

Teagueb: /* Calibration Experiment 1 Results */

2022-05-03T19:45:44Z

‎Calibration Experiment 1 Results

@@ Line 96: / Line 96: @@
 ==Knock-out Protocol==
-  '''''Safety Note''''': When conducting any experiment it's important to wear the proper PPE for yourself as well as to protect your experiments in our lab we wore a lab coat and gloves. While we were in our cleanroom doing the UV Light Exposure we wore additional equipment including hairnets, coverall top, boot covers, and gloves.
+  '''''Safety Note''''': When conducting any experiment, it's essential to wear the proper PPE for yourself and protect your experiments. In our lab, we wore lab coats and gloves. While we were in our cleanroom doing the UV Light Exposure, we wore additional equipment, including hairnets, coverall top, boot covers, and gloves.
 #Calculate the amount of pure cell culture and amount of PBS in order to get 200 cells per mL
@@ Line 108: / Line 108: @@
 #Turn on the UV Light, set it to 400 Watts
 #Remove the top off of the cell culture plate
-# Expose the single-cell plate for 600 seconds under the UV Light, keeping the control away from the light
+# Expose the single-cell plate for 600 seconds under the UV light, keeping the control away from the light
 #Repeat Steps 8-10 until finished
 #Place cell culture plate in 30 degrees C incubator for 48 hours

@@ Line 108: / Line 108: @@
 #Turn on the UV Light, set it to 400 Watts
 #Remove the top off of the cell culture plate
-# Expose the single-cell plate for 600 seconds under the UV Light
+# Expose the single-cell plate for 600 seconds under the UV Light, keeping the control away from the light
 #Repeat Steps 8-10 until finished
 #Place cell culture plate in 30 degrees C incubator for 48 hours

@@ Line 105: / Line 105: @@
 #Pipette the full 500uL of the PBS and cell mixture into each 60 mm cell culture plate with agarose in it. This one will not be exposed to UV light and will be designated as the control
 #Add Glass beads and shake for 30 seconds, then remove glass beads
 #Turn on the UV Light, set it to 400 Watts
 #Remove the top off of the cell culture plate
 # Expose the single-cell plate for 600 seconds under the UV Light

@@ Line 35: / Line 35: @@
 #Turn on the UV Light, set it to 400 Watts
 #Label cell culture with interval time (1 Second, 2 Seconds, etc.)
-#Remove the top off the cell culture plate
+#Remove the top off of the cell culture plate
 # Expose the single-cell plate for the designated time under the UV Light
 #Repeat Steps 8-10 until out of intervals.
@@ Line 42: / Line 42: @@
 ===Calibration Experiment 2===
-Due to the results of calibration 1 the time gaps left us needing to do another experiment to test the time between 500 seconds and 1000 seconds. We did new time intervals of 550 seconds, 600 seconds, 650 seconds, 700 seconds, 750 seconds, 800 seconds, 850 seconds, and 950 seconds.
+Due to the results of calibration 1, the time gaps left us needing to do another experiment to test the time between 500 seconds and 1000 seconds. We did new time intervals of 550 seconds, 600 seconds, 650 seconds, 700 seconds, 750 seconds, 800 seconds, 850 seconds, and 950 seconds.
 #Calculate the amount of pure cell culture and amount of PBS in order to get 1000 cells per mL
@@ Line 52: / Line 52: @@
 #Turn on the UV Light, set it to 400 Watts
 #Label cell culture with interval time (550 Seconds,600 Seconds, etc.)
-#Remove the top off the cell culture plate
+#Remove the top off of the cell culture plate
 # Expose the single-cell plate for a designated time under the UV Light
 #Repeat Steps 8-10 until out of intervals.
@@ Line 102: / Line 102: @@
 #Using a P-1000 Micropipette, pipette the amount of PBS calculated into the same micropipette tube as step 2
 #Mix the PBS and cell culture either by hand or by using a vortexer
-#Pipette 500uL of the PBS and cell mixture into each 60 mm cell culture plate with agarose in it.
+#Pipette the full 500uL of the PBS and cell mixture into each 60 mm cell culture plate with agarose in it.
-#Pipette 500uL of the PBS and cell mixture into each 60 mm cell culture plate with agarose in it. This one will not be exposed to UV light and will be designated as the control
+#Pipette the full 500uL of the PBS and cell mixture into each 60 mm cell culture plate with agarose in it. This one will not be exposed to UV light and will be designated as the control
 #Add Glass beads and shake for 30 seconds, then remove glass beads
 #Turn on the UV Light, set it to 400 Watts
 #Label cell culture with the Gene Number
-#Remove the top off the cell culture plate
+#Remove the top off of the cell culture plate
 # Expose the single-cell plate for 600 seconds under the UV Light
 #Repeat Steps 8-10 until finished

@@ Line 102: / Line 102: @@
 #Using a P-1000 Micropipette, pipette the amount of PBS calculated into the same micropipette tube as step 2
 #Mix the PBS and cell culture either by hand or by using a vortexer
-#Pipette the full 500uL of the PBS and cell mixture into each 60 mm cell culture plate with agarose in it.
+#Pipette 500uL of the PBS and cell mixture into each 60 mm cell culture plate with agarose in it.
-#Pipette the full 500uL of the PBS and cell mixture into each 60 mm cell culture plate with agarose in it. This one will not be exposed to UV light and will be designated as the control
+#Pipette 500uL of the PBS and cell mixture into each 60 mm cell culture plate with agarose in it. This one will not be exposed to UV light and will be designated as the control
 #Add Glass beads and shake for 30 seconds, then remove glass beads
 #Turn on the UV Light, set it to 400 Watts

@@ Line 14: / Line 14: @@
 *Fine Tip Sharpie
 *60mm Culture Dish
 *Vortexer
 ==Calibration Protocol==

@@ Line 60: / Line 60: @@
 '''Results'''
 Second
-[[File:Ashton1Sec.jpg|100px|]]
+[[File:Ashton1Sec.jpg|200px|]]
 Seconds
-[[File:Ashton500Sec.jpg|100px|]]
+[[File:Ashton500Sec.jpg|200px|]]
 Seconds
-[[File:Ashton1000Sec.jpg|100px|]]
+[[File:Ashton1000Sec.jpg|200px|]]