Agarose gel electrophoresis

Directions for the nano drop was run accordingly to machine directions. For electrophoresis, 10 µl of the sample was transferred into a clean microfuge tube with 2 µl of 6x loading dye. The portion of the sample was pipetted into an agarose gel and run at 120V for roughly 60minutes. Upon completion the gel was photographed for conformation. PCR Preparation A master mix was prepared in accordance to the PCR set up table provided by laboratory…

Gel electrophoresis allows the products of the PCR reactions to be analyzed in order to determine the identity of the tetracycline resistance plasmid. First, in order to see the DNA fragments, move across the gel, add 2 uL of bromopenol blue tracking dye to each of the tubes. In the first well add 10 uL of the PCR DNA ladder as a comparison. Carefully load 15 uL from each of the samples into the wells following the chart below. After the samples are loaded, place the lid onto the gel rig,…

examined the other two loci, but gained limited data from them. To analyze the seal samples, the researchers extracted DNA from the samples and amplified the short tandem repeats using PCR. Then, the scientists ran the PCR products through agarose gel electrophoresis stained with ethidium bromide. The researchers measured the sizes of the bands by comparing them to standard fragments with known sizes. After the completion of the process, the scientists compared the seal’s genetic similarity by…

The gels are made of agarose. To begin, we used .48 grams of agarose to prepare 30µl of a 1.6% agarose solution. We then transferred the weighted agarose to a 125-ml Erlenmeyer flask and added 30 ml of 1X TBE buffer. The agarose was then heated in a microwave for 30 seconds, before adding ethidium bromide which will stain the DNA fragments as they move through the gel. After mixing the agarose and bromide, we constructed the chamber and proceeded to carefully pour the agarose into the chamber…

pKN800 plasmid DNA. The plasmid was purified and isolated, then cut with PstI restriction enzyme to create a sample of PstI-cut and uncut pKN800 plasmid DNA and to distinguish orientation A from B. These samples were loaded onto a 0.8% agarose gel for electrophoresis. Both cut and uncut plasmid DNA were transformed into E. coli DH5α. Finally, the transformed DNA was plated on LB and LB with ampicillin agar plates. The procedure for this experiment was followed on pg. 16-23 of the MB 311…

experiment, agarose gel electrophoresis was used in to analysis the DNA that was extracted. The nucleic acid is at PH of 8 which allows the material to move down the electric field. The nucleic acid moves from positive to negative electric field. The ETBR is used to make the nucleic acid visible in the fluorescent light and the binding to the DNA. The “stop buffer” which is EDTA and bromophenol blue. The Bromophenol helped make the progress of making the DNA visible when it is injected in the…

Each tubes were incubated for 60 minutes in a 37oC water bath to guarantee cleavage of DNA. Eventually, 5 µl of 10x gel loading solution was added to each reaction in order to stop the reactions. Then each solutions was mixed again then soon placed in heated bath for two minutes, eventually added to the electrophoresis gel for about 60 minutes. The .8% agarose gel used for electrophoresis aided in slowing the migration of the DNA fragments since large DNA fragments migrate faster than expected…

Introduction In the United States a suspect in a crime is innocent until they are proven in court of law to be guilty beyond doubt(Cornell web), for now at least. This need to prove guilt in a suspect means the use of DNA to determine if the suspect was at least present at the crime scene is a powerful tool because no one person can change their DNA at will. The use of DNA as way to identify people has it 's origins in the United Kingdom because of a technique created by Dr. Alex Jeffreys used…

Electrophoresis will be carried out in basic medium, composed of a TBE IX buffer (0,89M Tris, 0.8M Boique acid, 0.02 mM EDTA; pH 8.4 disodium). The nucleic acids, polyanionic macromolecules uniformly negativemen loads, when placed in a constant electric field will then migrate through the gel towards the anode. The speed of movement of the fragments through the gel mesh will therefore vary depending on the concetration in agarose gel, but also the molecular weight (number…

of thermal cycle and repeat. Each DNA used in PCR is serve as a template. The PCR product then is analyzed by agarose gel electrophoresis. Agarose gel electrophoresis is the common and easy way to analyze DNA where that DNA is visualized in the gel. There is a problem may be occur during PCR reaction and electrophoresis such as DNA product appear variable in non-specific on agarose gels. Sometimes no product form at all. (Lorenz,2012). Other is mutation occur during amplication that result in a…