Design a plan for collecting data to show that all biological systems are affected by complex biotic and abiotic interactions.
Use models to predict and justify that changes in the subcomponents of a biological polymer affect the functionality of the molecule.
Analyze data to identify how molecular interactions affect structure and function.
Everything will be in the lab notebook. DO NOT WRITE ON A SEPARATE SHEET OF PAPER.
I will give you the semi-log graph paper and the standard curve to refer to.
You will be working in groups of 3 (and 1 pair) = 4 groups total
This will be done over THREE (3) days due to state testing. Due to this, you need to be on point as we don't have any wiggle room for error.
Things might not work the way they're supposed to but that's OK. It's science; things don't always have to work out!
Obtain a gel-casting tray. Put the dams into the slots at each end (or masking tape).
Insert the small side of the comb into a slot on either end of the casting tray.
The melted agarose gel should be cooled to 55℃. Using heat resistant gloves, pour the agarose gel into your casting tray, so the depth is 1/4 in and covers the comb.
Let the gel solidify until it is opaque and whitish in appearance. (~30 mins)
Remove your gels, put them in a ziplock bag with buffer solution, and store in the refrigerator until next class.
Obtain your gel and place it into a electrophoresis chamber.
Pour running buffer into the chamber so it covers your gel.
Using a micropipette, draw up 10µL of Sample #1 (uncut Bacteriophage DNA).
Put the DNA into well 1 by holding the tip just inside the well. Be careful not to puncture the bottom of the well or the sample will leak.
Repeat steps 3 & 4, using a new pipette tip each time, for the remaining sample into their corresponding wells:
Well 2: DNA/EcoRI digest
Well 3: DNA/HindIII digest
Well 4: EcoRI/HindIII digest
Place the lid on the chamber, matching red and black ends. Connect plugs into their appropriate spots on the power supply. Use 150 V setting.
Turn on the power supply and let it run for ~30 min (or until the dye is 1cm from the end of the gel).
Turn off the power supply, unplug connection, and open lid.
Place the gel in a staining solution in a ziplock bag. This will sit overnight and in the refrigerator until our next class.
Get your gel. You may have to rinse off the stain under warm water.
Measure the distance each band migrated (in mm) in each lane from the bottom of the well to the top of the band.
Record your results in the data table.
Create an accurate drawing of your gel, indicating the position of the DNA bands in each sample.
Create a graph and answer questions.