DNA lab 2 (temporary): Agarose Gel Electrophoresis How to pour, load, and run an agarose gel. MATERIALS Buffers and Solutions Agarose solutions (please see Step 3) DNA staining solution Electrophoresis buffer 6x Gel-loading buffer Nucleic Acids and Oligonucleotides DNA samples DNA size standards Samples of DNAs of known size are typically generated by restriction enzyme digestion of a plasmid or bacteriophage DNA of known sequence. Alternatively, they are produced by ligating a monomer
The research work was undertaken with the aim of study Agrolistic transformation in Sugarcane and studies of associated problems. The work was carried out in the Department of Molecular Biology and Genetic Engineering, Vasantdada Sugar Institute, Pune during Jan 2014- May 2014. The materials used and the methods adopted are presented below: Plant material used Top portion of sugarcane of age varying from 4-10 months is used as initial explants however sugarcane of more than 6 months was not preferred
In the preparation of agarose gel, agarose powder will be mixed with buffer. Agarose powders need to be weighed first before mixing it with buffer. When 1 gram agarose gel is added, it will be followed by 100 ml of buffer. The ratio is 1:100 for agarose powder and buffer respectively. Then, the solution of agarose powder and buffer is put in the microwave in order to melt the agarose until the solution become clear (Carson & Robertson, 2005). Agarose gel is poured in the casting trays. Actually casting
the region of DNA. In the taster gene, after amplification, a restriction digest was performed to differentiate between the two alleles. The digest was able to show differentiation because those with the “T” allele would have two bands from gel electrophoresis and those with “t” will have one band because the restriction enzyme doesn’t cut it. For the PV92, we were able to distinguish between the alleles due to the added length of the Alu element. Those... ... middle of paper ... ...G.E., Ioannou
Then DNA ladder and flow cytometry assays were done for detection of apoptosis in NIH-3T3 cell line. RESULT: Primary and late apoptosis in the treated cells was determined by flow cytometry analysis. Accordingly DNA ladder assay using 1.5 % gel electrophoresis showed fragmentation in the DNA of treated cells. CONCLUSSION: This research provides a fast, simple and cost-effective method in observing apoptosis in mammalian cells and could be use in cancer research and genotoxicity. Introduction Apoptosis
DNA. Most of the time, this is done using a technique known as gel electrophoresis. Gel electrophoresis is a method used to separate the macromolecules that make up nucleic acids, such as DNA and RNA, along with proteins. Gel electrophoresis is significant because it has given scientists insight on what cells cause certain diseases and has led to advancements in DNA and fingerprint identification. My experiment will use gel electrophoresis to compare samples of natural and synthetic food dyes. The background
consisted of three parts. In the first part, plasmid DNA was incorporated into bacterial cells. The second part consisted observing the new phenotypic traits on agarose plates, and isolating the plasmid DNA from the transformed bacterial cells to be used in PCR reactions. The final part was analyzing the PCR reactions on agarose DNA gel electrophoresis. Materials and methods: Part 1 - Bioluminescence Materials: • sharpie • 37 oC water bath • Ice • Sterile transfer pipette • Foam tube rack • Transformation
This causes the DNA fragments to move through the gel depending on their sizes. With this, the DNA fragments will show a sample that will determine how large they are to one another. Gel electrophoresis uses a horizontal gel-like slab. These gels are made of polysaccharide called agarose, which is dry, powdered flakes. When the agarose is heated in a buffer, it makes the gel form solid, slightly squishy gels. (Dickey, J. L. 2012) At one end of the gel, there is square shaped space that is called wells
Chloroplast fractionation: Nucleic acid and protein analysis via gel electrophoresis ABSTRACT: Chloroplasts carry out photosynthetic processes to meet the metabolic demands of plant cells (Alberts, 2008). They consist of an inner thylakoid membrane and a stroma. (Parent et. al, 2008).In this experiment we demonstrate the unique protein compositions of isolated thylakoid and stromal fractions from broken and whole spinach chloroplasts. Because these compartments carry out different metabolic processes
Thanks to TV shows like CSI, many people are familiar with the use of gel electrophoresis to separate macromolecules like DNA. However, gel electrophoresis can also be used to separate out proteins. Different proteins have different sizes, mainly due to the number of amino acid building blocks in their structure. Chemical modifications attached to the protein also affect its size. Different proteins also have different charges. This can result from both the types of amino acid used to construct
DNA Fingerprinting Using Agarose Gel Electrophoresis Introduction Agarose gel electrophoresis is a form of gel electrophoresis that can separate a mix of DNA and proteins through agarose gel. It separates DNA by length or size through gel when an electric current is applied. Shorter fragments travel faster than long through the gel allowing for matches to be identified by similarity. The fragment length of DNA is different for each individual because sequences cut at specific sites. PCR or polymerase
Electrophoresis is an analytical technique for the analysis of macromolecules like proteins and nucleic acids. This technique was discovered and first used in 1937 by a Swedish biochemist Arne Tiselius . The electrophoretic effect is based on the theory of Debye - Huckel - Onsager where this theory of electrolytic dissociation accept the fact that charged particles move up under the influence of electrostatic forces to an electrode of opposite charge is applied when a potential difference in a solution
Introduction Alu elements are a class of transposable genes found exclusively in the genomic sequences of primates. Averaging in lengths of approximately 300 base pairs, Alu elements are classified as being short interspersed elements, more commonly referred to by the acronym SINEs. These elements interject themselves into the DNA sequence by means of retroposition. Once established into the genome, Alu elements are considered to be stable, only rarely being subjected to deletion. Initial studies
investigate the enamel proteins in various types of amelogenesis imperfecta and to fully deduce if amelogenin was retained in the fully developed amelogenesis imperfect enamel. The primary Biochemical method used in this study was the SDS-Page Electrophoresis ... ... middle of paper ... ...ed C, J. W. (1992). Enamel Protein in the different type sof Amelogenesis Imperfecta. The chemistry and biology of mineralized Tissues , 441-450. C, W. (1989). Amelogenesis Imperfecta dentinogenesis imperfecta
the cations. And here A- will be negatively charged which is masked by positively charged molecule B+. Mobile phase will have cations (say C+) which need to be separate. As the idea here is the exchange of ion. Negative part (A-) attached to the agarose is already attached to positive part (B+) so to remove B+ we need to add another negative part(D-) . It will help to remove B+ and in addition of C+ to A-. Then this desired cation C+ will be separated by Buffer solution. By using cation exchange
3/11/16 Introduction The aim of this experiment is to separate the protein samples based on their molecular size using the SDS-PAGE technique and to detect EGFP protein by carrying out a western blot. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) is a technique used in the lab for the separation of proteins by their molecular weight. SDS is a detergent used in PAGE because its main role is to break down the disulphide bonds which disrupts the tertiary structure of the proteins
Protein extractions from unidentified fish samples were separated according to the molecular weights by SDS polyacrylamide gel electrophoresis. Since some of these proteins are shared between fishes, phylogenetic evaluation was reached. Western blot analysis was used to identify four unknown species of aquatic animals via comparison of actin/myosin bands. According to the results of this assay, the best estimate is that the unidentified aquatic animals are specimens of salmon, tilapia, cod, and shrimp
crude lysate, it is possible that proteins are in very small quantities and thus could not be expressed definitively. The source of this error was more likely in the beginning of the lab, but the error cannot be traced with certainty as the original gels for ligations, transformations and digests were penetrated and could not yield at a discernable
determined to have an isoelectric point of 3.5 to 5 and will be analysed to ensure this result by use of an isoelectric focusing gel. This will establish whether the in process sample analyzed shows any potential for protein modification such as glycosylation or phosphorylation and following this run will be measured for the correct isoelectric point by use of 2D gel electrophoresis. (LifeTechnologies, 2014)
until the dyes reach the end of the gel. After electrophoresis, use Ethidium Bromide (C21H20BrN3), which links with DNA molecules and fluoresces under ultraviolet (UV) light to observe the DNA fragments on the gel. Photographing the lit gel under ultraviolet light in a dark room to record the result. Movement speed of DNA in the electric field depends on many factors such as electric power, buffer composition, concentration agarose gels. The higher concentrations of gel, the stronger resistance, which