Protocol 4 - Gel Electrophoresis

there are many components of the lanza system that first need to be assembled you should be in possession of the following components a camera hood a power supply a gel box a laptop and lots of cores red and black cords hardwired to the gel box will be plugged into the power supply eventually but you should only do so right before running the gel on each of the cords a label will be provided to help orient you to their connectivity this cord connects the gel box to a wall power outlet this cord connects the power supply to a wall power outlet on the inside of the laptop you will find a username and password to login with once the computer is powered on to power on the laptop must be plugged into a wall power outlet as the battery on these repurposed laptops has little to no life the camera hood should be plugged directly into a USB port on the laptop and does not require an extra power supply once everything is plugged in it is advisable to turn everything on to ensure that it is working before beginning the experiment if everything is working properly then you can set up the precast gel a gel that is appropriate for your class size and sample needs will be given to you simply remove the packaging from the gel cassette the cassette should slide into the gel box with an affirming click and be flush with the outer edges of the gel box before loading the gel with samples the wells must be washed with water it is easiest to use the squirt bottle provided pour out the excess water and dab the gel with a kimwipe or paper towel before returning it to the gel box all students should use the same gel to visualize their samples in this example the for students that amplified actin and three will run their samples with five other classmates with one negative control from one of the groups additionally a DNA ladder has been provided for visual size comparison all of these solutions should be red or purple indicating that they have been mixed with a loading dye to allow for visualization and settling into the wells once again mix and collect all samples before proceeding load three microliters of a sample into its designated well the diagram provided at the end of this protocol should serve as a template for loading skip the first well of the gel as this is where the latter will be loaded but we'll be done so last take your time to orient the tip into the second well the tip should not be jammed to the bottom of the well nor hover above the well but just on the inside of the plastic rim it is fine to use your other hand to stabilize your pipetting wrist expel your sample slowly you want to avoid bubbles or flow out of the well you should be able to see the colored ID fill the well once you started loading the gel the gel should not be taken out of the doc or it may cause spillover between the samples repeat this process for all remaining samples add three microliters of the ladder to the first well of each row in the same manner press the Volt button to ensure that your power supply is running at 275 volts then press the time button and navigate using the arrows to the desired run length in this case a one tear gel needs to have electrical current for four minutes plug the red and black cords into the respective red and black receptors on the power supply there are two pairs of these receptors plug the cords into one pair do not split them amongst the pair's to see the DNA bands migrating through the gel in real time press the light button on the gel dock this blue light though bright is not harmful to the eyes press the Run button to begin electrophoresis it will take a minute or so before the bands start to be visible once the timer has beat or the bands have migrated halfway down the gel press the stop button turn off the power supply and unplug the bread and black cords to capture an image of the gel leave the blue light of the gel box on and double-click on the flash gel icon on the computer when you mount the camera hood to the gel box an image of your gel should appear on the screen press the camera icon to capture the image and save the image on the computer in a recognizable location with a recognizable name it is fine for you to transfer this image to your own storage device from this laptop

Channel: The Jackson Laboratory

Views: 45,697

Rating: 4.891892 out of 5

Keywords: dna, sequencing, genetics, genomics

Id: AK8A5OREk34

Length: 10min 45sec (645 seconds)

Published: Thu Jul 02 2015