StudentShare solutions
Triangle menu

DNA Restriction and Electrophoresis - Lab Report Example

Not dowloaded yet

Extract of sample
DNA Restriction and Electrophoresis

The various endonucleases used areECORI, Bain HI, and Hind III.Each of these enzymes has at least five or more restriction sites on the chromosome and it consequently produces six or more restriction fragments of different lengths.
The DNA strands are then immersed in a well of 0.8% agarose gel, which is the chosen medium.An electrical field, is applied to the gel evenly which makes the DNA fragments move faster from their original placement towards the positively charged electrode. The smaller DNA fragments travel faster through the gel than the larger ones. The gel actually acts like a sieve. Depending on the restriction enzyme used the DNA fragments separate into distinct bands during electrophoresis. The bands are then stained with a dye to make the invidual restriction enzymes evidently distinguishing them with a dye that attaches itself to the DNA molecule.
Electrophoresing can be detected by hydrogen gas bubbling from the negative electrode and oxygen being given off by the positive electrode. These are products of the electrolysis of water. This is the first sign that current is flowing through electrophoresis system. Shortly afterwards bands of loading dye should be seen moving into the gel and migrating towards the positive pole of the apparatus. ...
The movement of the DNA fragments are highlighted by the dye which forms coloured bands.(See above and below)

Results: Depending on the endonuclease used, the digested DNA samples, exhibit the characteristic bands produced by each restriction. The enzyme is seen because of a dye that is introduced and shows the pattern of movement of the DNA fragments.

Various restriction enzymes: There are several types of restriction enzymes, each of which react differently with the DNA and create different cuts. there are some that cut a three base pair sequence while others can cut four six or even eight. Each enzyme has different properties that are factors that determine the efficiency with which the DNA is cut and under what conditions optimum results will be obtained. Manufacturers of these restriction enzymes usually provide a specific buffer solution that is unique to the enzyme. This buffer solution is a mix of cat ions and other components that optimize the efficiency of the enzyme for cutting. Different restriction enzymes have different optimal temperatures under which they function.

Conclusion: It can be observed that each sample of purified DNA reacts differently with the varied restriction enzymes. The fourth sample used for the negative control remains intact; as it has been incubated without, an endonuclease.The new DNA fragment can then be extracted from the gel by cutting it and doing gel purification. Vector ligation is used as the insert point. The ligation of the vector has to be cut also by a restriction ...Show more


Purpose: Electrophoresis is a method used to separate DNA molecules by sized restrictions are the enzymes used to prepare DNA for analysis or other manipulations. Here we examine the genotypic analysis of DNA using restriction enzymes and gel electrophoresis.
Author : reillykody
DNA Restriction and Electrophoresis essay example
Read Text Preview
Save Your Time for More Important Things
Let us write or edit the lab report on your topic
"DNA Restriction and Electrophoresis"
with a personal 20% discount.
Grab the best paper

Related Essays

Practical 1: Aim: To perform plasmid miniprep to isolate pBlueSkript KS II (+) from E.coli and to perform double restriction digestion on pProEx with the enzymes BamHI and Hind III to release the FtsZ insert. Secondly, to increase the efficiency of ligation by alkaline phosphatase treatment of the linearised plasmid.
22 pages (5500 words) Lab Report
Expression Of Recombinant Tick Histamine-Binding Protein
EXPRESSION OF RECOMBINANT TICK HISTAMINE-BINDING PROTEIN TC11485 FROM Rhipicephalus microplus USING PPICZ-ALPHA IN Pichia pastoris Abstract Histamine-binding proteins (HBP) are ubiquitous biomolecules with pharmacologic importance, since they have an immunosuppressing activity, which can help prevent adverse health conditions such as histamine shock.
30 pages (7500 words) Lab Report
Agarose Gel Electrophoresis of DNA
(Westermeier 2006). Agarose gel electrophoresis is the most common and easiest technique to separate the DNA fragments. Agarose is a polymer that forms helical linking strands between the molecules and they are held together by hydrogen bonds. Based on the concentration of the agarose in the gel, the pore size also varies.
5 pages (1250 words) Lab Report
Protein Extraction and Gel Electrophoresis
Proteomic is generally defined as the direct analysis of proteins in terms of their presence and relative abundance. Gel electrophoresis is a significant methodology employed for extraction of proteins in proteome analysis. The most commonly used technique in gel electrophoresis is Sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
5 pages (1250 words) Lab Report
Lab Report
Since 19th century scientists were aware of some unique identity of an individual which distinguish from others, like finger prints. In 1970, DNA was established as key element in human life and which makes individual as unique creature. Variability in DNA sequences among individual to individual have attracted forensic experts to develop technique based on DNA to identify individual or criminal similar to finger prints.
9 pages (2250 words) Lab Report
Biochemistry Lab Report
To confirm the success of transformation, the recombinant DNA was isolated and purified from the transformants followed by digestion using HIND III /EcoRV. The GFP protein was then purified from the transformants using hydrophobic interaction chromatography (HIC) and its purity confirmed using immunoblotting and SDS-Page.
3 pages (750 words) Lab Report
Molecular Biology
Abbreviations: Inr= initiator sequence of the RNA polymerase II promoter; UCE= upstream control element of the RNA polymerase I promoter. A 29 kilo base (kb) linear DNA fragment is digested with ApeK I, Bst I and Cla I, individually and in combination, and the resulting DNA fragment sizes are determined by agarose gel electrophoresis.
7 pages (1750 words) Lab Report
DNA Practical
The DNA from mitochondria and chloroplast is responsible for cytoplasmic inheritance or maternal inheritance. DNA is a double stranded nucleic acid and both the strands run antiparallel. It is made up of pentose sugar (de-oxy ribose), phosphate group and nitrogen base pairs (Double ringed Purines: Adenine and Guanine and Single ringed Pyrimidine: Cytosine and Thymine).
4 pages (1000 words) Lab Report
Gene technology
The insert (16s rDNA = 1.5 kb; plasmid = 3.5 kb). The band on our slide is dark and broad therefore exact size determination is difficult. However, the size is slightly more than 4.0 kb, which is near the expected
3 pages (750 words) Lab Report
Lab report introduction
The mutant gene which we shall be testing does not have a restriction site for the restriction enzyme HindIII that we shall use in the practical. The wild type gene on the other hand has a restriction site and so HindIII will bind for activity to
1 pages (250 words) Lab Report
Get a custom paper written
by a pro under your requirements!
Win a special DISCOUNT!
Put in your e-mail and click the button with your lucky finger
Your email
Comments (0)
Rate this paper:
Thank you! Your comment has been sent and will be posted after moderation