Gel electrophoresis

Background DNA molecules can be too big to analyze through electrophoresis, some they must be cut up into smaller pieces. In order to cleave dsDNA at the specific sites, making sure each fragment contain the desired sequence, restriction enzymes are used. Restriction enzymes can recognize a specific sequence of nucleotide and hydrolyze the bond of the DNA backbone. Hence, the fragments from the same DNA source will also be the same if cleave by the same restriction enzymes. Restriction enzymes…

Gel electrophoresis is a method used for separation and analysis of molecules such as DNA, RNA, and proteins, based on their sizes and polarity. DNA (deoxyribonucleic acid) is a molecule that carries most of our genetic information, and possesses a negative charge. During gel electrophoresis, DNA fragments can migrate through the gel also known as agarose when placed in a powerful electrical field. The rate at which the DNA fragments will move through the gel depends on their relative size.…

using agarose gel electrophoresis. Species identification was performed by importing finalized CO1 sequences into two DNA databases to ensure accurate identification. Phylogenic analysis was then conducted to determine relationships between identified species. Electrophoresis Electrophoresis of DNA run in 2% agarose gel showed successful extraction of DNA in samples 1-8, 12 and 14-16. Samples were of good quality with high molecular weights (Figure 2). Samples 11 and 13 in Row 1 of gel 2 were…

As DNA is a negatively charged molecule the samples when run through the gel move towards the positive end of the gel, this is important to remember during the setup of the gel so that the electrodes are placed correctly. For each region of DNA under investigation the size of the molecule depends on the amount of nucleotides within that sequence, for the first experiment this…

on the SDS-PAGE gel than the Simple-PAGE gel. This is due to the fact that SDS denatures proteins, allowing them to be unfolded and individual polypeptide chains. The Simple-PAGE gel does not denature the proteins, and thus proteins are run in a folded and potentially active shape. An example of this is that alkaline phosphatase activity was detected in the Sigma Red stained Simple-PAGE gel but nor the SDS-PAGE gel. Thus AP had the proper folding to be active in the Simple-PAGE gel. Based on…

In this experiment Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) was performed under reducing conditions to estimate the molecular weight of proteins within a sample. The protein sample was from the previous using anionic exchange chromatography to partially purify N-acetyl--D-hexosaminidase, Lab 3. The molecular weight was determined through the interpolation of a standard curve of proteins with a known molecular weight. The molecular weight of band 1 was determined to…

Phase 2 – Agarose Gel Electrophoresis 1. Obtain our four H, E, L, and P labeled micro tubes and place them on ice. 2. Set the micro pipet to 2.0 ul. 3. Transfer 2.0 ul blue dye to each of the four H, E, L, and P micro tubes. 4. Mix the four micro tubes by using this two-step method: * Place each tube briefly on the vortex to get the components to collect at the bottom of the micro tubes * Pulse spin the four micro tubes in the centrifuge to mix all of the components completely. 5.…

Analytical agarose gel electrophoresis This molecular tool is mainly used to separate and identify DNA fragments and in some cases also to purify certain DNA molecules that can be cut out of the gel after the electrophoresis. DNA molecules are negatively charged, and by loading them into an agarose gel and applying a current creating a uniform electrical field, these molecules will move across the gel. How far they travel, depends on their electrophoretic mobility. Size, structure and total…

just the DNA in its membrane. Later some Elution Buffer was used to separate the DNA from the membrane of the spin column. Day two consisted of making the plasmid DNA resistant to Kanamycin and Ampicillin. Day three we made 0.8% agarose gel and ran electrophoresis to be able to see the weight of the DNA. Day four we genetically transferred the ligated plasmid DNA to E.coli and plated the cell mixture. Day five we looked at our results and went over them. All of this works together for bacterial…

smaller parts. The fragments of DNA are then applied to a jelly-like substance called agarose gel, and an electric current is then sent through the gel. Because the strands of DNA are negatively charged, they will move across the surface of the gel, and the smaller pieces will move farther. The gel is hard to handle, so it is covered in thin nylon membrane, and a layer of paper towels that draws moisture from the gel. As the moisture is being transferred to the paper towel, the DNA is being…