Agarose gel electrophoresis

combined into one solution. The resulting solution will then be subjected to 2-dimensional gel electrophoresis. This will be done by transferring the mixed protein solution to isoelectric tube gel (pH 3-10) and subjecting it to a constant electric current (145mA). After the gel has been allowed to run for about an hour, the isoelectric tube will be rinsed with running buffer and set aside. Next, an SDS gel will be prepared in order to separate the proteins based on mass. This will be done by…

mixture of three molecules—blue dextran, cytochrome c, and potassium chromate—through the use of gel-filtration chromatography. The volume at which each molecule elutes will be recorded in order to determine the accuracy of separation. The purpose of Experiment 5-2 is to purify the E. coli enzyme, β-Galactosidase through the use of affinity chromatography on P-aminobenzyl-1-thio-β-D-galactopyranoside agarose, which is a good substrate for the enzyme. This enzyme will be eluted at high pH. The…

DNA to come inside and interact with the bacterial DNA. The DNA gel electrophoresis showed that there were no fragments…

In this experiment, a SDS-PAGE gel was used to analyze the protein samples from the MBP-AP and WT-AP experiments. The samples are then referenced to the ladder to determine the molecular weight of the MBP-AP and WT-AP proteins. Then the UV absorbance of the two proteins from 240 nm to 340 nm is determined using a nanovolume cuvette. The absorbance at 280 nm was then used in conjunction with data from previous experiments to determine the concentration of the MBP-AP and WT-AP protein samples.…

Bovine serum albumin (BSA) is usually used for generation of nanoparticles (NPs) in drug delivery systems. This paper presents a study on the simple coacervation method for fabrication of BSA nanoparticles.An artificial neural network (ANN) model was developed to predict the diameter of nanoparticles based on experimental data. Effect of pH, temperature, initial BSA and cross linker concentration, molar ratio of BSA to ethanol, agitation rate and rate of ethanol addition were investigated on the…

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…

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…

be analyzed from a variety of human samples including blood, semen, saliva, urine, hair, buccal (cheek cells), tissues, or bones. The polymerase chain reaction (PCR) is used to amplify the genomic DNA from a sample and electrophoresis is used to arrange the segment.…

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…

honeybees. The hypothesis is that the four unknown bees are hybridized with a European mother and an African father. The wing-lengths of European, African, and unknown bees were measured, and the mtDNA of eight different bees were subjected to gel electrophoresis. The results showed that African bees were small with one band of mtDNA, European bees were large with two bands of mtDNA, and the unknown bees were of various sizes with two bands of mtDNA. The hypothesis was supported due to the…