Bacteria Growth in Different Test Tubes
The purpose of this lab is to learn about restriction enzymes, plasmids, and cloning genes. In class, we learned a lot about the structure of DNA and genes, which were both very helpful while performing this experiment. Restriction enzymes have the ability to cut DNA at specific sequences. Plasmids are perfect for genetic engineering because they can replicate and initiate transcription. Performing this experiment helped us learn more about the process of cloning a gene. We also learned about transcription and translation in class, which are both crucial parts in the process of cloning a gene. It was also necessary that we knew information about gel electrophoresis. In this process, DNA…show more content… Labeled two clean microfuge tubes “R+” and “R-”.
Added the following:
4.0 µL of 2.5xB to the R+ and R- tubes.
4.0 µL of RP to the R+ and R- tubes.
2.0 µL of RE to R+ tube.
2.0 µL of dH20 to R- tube.
Spun R+ and R- tubes in microcentrifuge for four seconds.
Placed tubes in 37o C water bath. Left in water bath for at least 60 minutes, then placed both of the tubes in freezer at -20o C.
Microfuge tube of non digested pARA-R from 2A (R-)
Microfuge tube of digested pARA-R from 2A (R+)
Microfuge tube of loading dye (LD)
Microfuge tube of DNA ladder (M)
Equipment and Supplies
Disposable gloves and lab goggles
Tip box of disposable pipette tips
Electrophoresis box with 0.8% agarose gel
Waste container (for used tips and microfuge tubes)
Obtained materials and checked for all reagents.
Added 2.0 µL of LD to R- and R+ tubes.
Spun R- and R+ tubes in microcentrifuge for several seconds.
Filled box with 1 xSB to level that just covers entire surface of gel.
Used a fresh pipette tip for each sample and dispensed 10.0 µL of DNA ladder (M), 10.0 µL of R-, and 10.0 µL of R+ into designated wells.
Plugged the well into a power supply.
Turned on power supply and set voltage to 130-135 V.
Let yellow LD run for about 40-50 minutes.