*Make 1% weight/volume Agarose gel in TAE buffer
*Make ttl volume of 50mL
*~0.5g/ 50 mL d H2O
*Warm solution for ~1 minute 40 seconds in microwave
*Place beaker solution into 55 degree C water bath for ~10 minutes
*Take boat of Electrophoresis apparatus and seal sides of boat with tape in order to place agar gel within
*Place comb (w/ "teeth") into empty boat along the side
*Place Agarose gel into boat to solidify
*Put a ttl of 10 microliters of restriction enzyme using a 10 microliter pipette into the 6 colored tubes w/ previously filled DNA samples
*Place the 6 DNA/enzyme samples into incubator at 37 degrees Celcius for 45 minutes
*Restriction enzymes: ECOR1 (E.coli restriction enzyme) and Pst1
*Each restriction enzyme cuts different sequences of bases
*Place the boat into the electrophoresis apparatus
*Take out comb and place colored tube samples of rest.enz/DNA samples into wells w/ 10 microliter micropipette
*1 well has DNA ladder (w/ known size fragments)
*1 well has "crime scene" sample (from colored tube sample labeled CS)
*Other 5 wells have "suspect" samples (from colored tube samples labeled 1-5)
*Add 5 microliters of blue dye to 6 colored tubes
*Remove tape from gel; put boat and gel into the electrophoresis apparatus
*Pour enough TAE buffer into the electrophoresis apparatus to cover the wells
*Place DNA ladder into one well, CS into another well, and colored tubes 1-5 into 5 different wells
*Be sure not to cross contaminate by getting sample/dye into other wells to be used by another sample/dye
*Attach Voltage cords to EC Apparatus and then to the electrophoresis apparatus
*Make sure wells are placed on the negative side of the electrophoresis apparatus b/c the DNA sample is negative and will then travel towards the positive side of the apparatus
*Set EC Apparatus at 120V on low range for 30 minutes
*Take gel out of boat and stain gel in FastBlast stain for 2-3 minutes
-Be careful when taking gel out because of the fragility where the wells are, it can easily be broken.
*Destain in warm tap water for ~15-20minutes changing out the H2O to quicken process to lighten up blue gel
-larger DNA will have harder time moving through the gel than smaller DNA.
-DNA is negatively charged and will go towards positive end
**Important Note- although this is a protocol for what was done in lab today, this is not a playbook for future experiments. The gel density, electrophoresis running time, amount of restriction enzymes and other factors will change with each experiment. **
