The final goal of this lab was to successfully measure the size of different Sam poles of DNA by placing each sample into a well in agrees gel and running a current t wrought a charged chamber. The DNA samples will move through the gel towards the p costive charge. Ideally, the DNA will move and create and sequence of smallest to largest. HTH s lab exposes us to DNA technology. Background Gel electrophoresis is used to separate macromolecules like DNA or RNA by is zee or proteins by charge.
To examine DNA and RNA, the fragments are placed in the e agrees wells and an electrical charge is sent through, pushing the negatively charged mole clues towards the positive side. The smaller the molecule, the less resistance it will face whew n hitting the pores of the gel, and the farther it will travel. Restriction enzymes are short nucleotide sequences used to cut DNA into SEG meets, separating the fragment into pieces. When cut, two different ends will be pro educed, a sticky end or a blunt end. When a sticky end is created, it makes the double helix stats greed, one end chills with an overhang above the other.
These ends can connect to an id enticed sequence cut by the same restriction enzymes or a very similar end. Blunt en DSL are created when a restriction enzyme cuts the double helix evenly. Materials One will need buffer solution, pipettes, an electrophoresis chamber, agrees, and three DNA samples consisting of an uncut sample, and a sample cut with Echo RI and one cut with Handbill to complete this lab. Methods TO Start things Off, the gel must be created. The mold has two open ends, there fore just be taped tightly and repetitively.
After pouring the agrees liquid into the e mold, it is mandatory that a ‘comb’ is placed in the mold to create the wells as the liquid solidifies. After 20 minutes, it has solidified, remove the comb and the tap and place the gel I n the chamber. The buffer solution is used to deliver the current to the agrees gel. Pour the buffer solution so it covers the gel. Add one of each sample of DNA to separate wells using a pipette. Cover the chamber and make sure the negative side of the circuit is on the same side as the wells.
After two hours of sitting in the electricity, remove the gel and stain it. Rinse t he gel thoroughly and let it sit in water for a day. Rest Its My petting must have been atrocious for only one of our three DNA sample s, the uncut sample, was visible. The uncut strand traveled two centimeters, forming g no bands as it was uncut. Discussion Our attempt at gel electrophoresis showed unbelievable potential, though I w loud describe it as feeble at best. In comparison with the lab manuals, our wimpy s ample conquered a whopping . CM less than the average uncut bacterial DNA We lacked results from the Score and Handbill samples altogether. The uncut sample was large, n o doubt, as it traveled 1/14th of the entire gel. Of the numerous places we could have made mistakes, there are three that w have created the most devastating end results. First off, we left Eleanor in chaw urge of sealing the ends of the mold and placing the comb.