We use cookies to create the best experience for you. Keep on browsing if you are OK with that, or find out how to manage cookies.
Nobody downloaded yet

Biochemistry - Lab Report Example

Comments (0)
The experiment aims at cloning and purification of the green fluorescent protein (GFP) protein using molecular techniques like transformation, restriction digestion and vertical gel electrophoresis. Briefly, the pGLO plasmid incorporated with the GFP gene, regulated by the arabinose inducible promoter was transformed into E.coli strain HB101…
Download full paper

Extract of sample

Download file to see previous pages... The gels were visualized by coomassie staining. The restriction digestion of recombinant DNA yielded the predicted band sizes upon electrophoresis, confirming the presence of the pGLO plasmid in the transformants. The presence of a single neat band at the expected size range upon immunoblotting indicates the successful isolation of the purified GFP protein.
Initially obtained from the jellyfish Aequorea victoria/Aequorea aequorea/Aequorea forskalea, the Green fluorescent protein (GFP) is composed of 238 amino acids with a molar mass of 26.9 kDa. Its typical three dimensional structure facilitates a specific set of cyclization reactions inside the protein giving rise to the tripeptide Ser65-Tyr66-Gly67 which forms the fluorophore, the fluorescent component of the protein, present on the alpha helix. This helix is actively shielded from the surrounding environment by the beta sheets, hence contributing to the use of the GFP gene as a reporter of gene expression or cellular protein localization.
In the present experiment, the GFP gene has been used to understand the mechanism of molecular cloning and subsequent protein purification procedures. The molecular cloning was carried out by transforming the pGLO plasmid vector with the GFP gene insert into the bacterial host E.coli strain HB101. ...
After appropriate incubation, the colonies were observed under normal light, followed by U.V light. Under normal light all the plates except plate with LB+Amp had growth
Lack of plasmid DNA on Plate with LB+Amp-pGLO might have rendered the colonies
susceptible to the antibiotic ampicillin, due to absence of genes required to deactivate ampicillin. Flourescence was observed in plates 3 and 4 under U.V light indicating the transformation of the plasmid DNA into the E.coli host. Plate 3, inspite of having ampicillin showed growth due to the presence of B-lactamase gene which can inactivate ampicillin. Plate 4 had the maximum number of colonies compared to the other plates.

This could be attributed to the addition of arabinose into the medium which selectively enhances the activity of the arabinose operon in which the GFP gene has been inserted, thus increasing the GFP protein which has the unique quality of fluorescence.
To confirm the insertion of the pGLO plasmid into the colonies on plates 3 and 4, the DNA from these colonies was purified using QAI kit method and subjected to restriction digestion using EcoRV and Hind III followed by electrophoresis, wherein an electric field was applied to the gel matrix. DNA molecules move towards the anode due to negativity of the charged phosphates along its backbone. The rate of migration of a particular DNA fragment is inversely proportional to its molecular weight; hence the fragments with the highest weight have the least mobility. Post electrophoresis, the ethidium bromide stained gel was visualized under UV light (Figure 1). A single restriction site specific to EcoRV is present on ...Download file to see next pagesRead More
Comments (0)
Click to create a comment
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
All the polymers are made of one or more of the twenty amino acids in different sequences and numbers. Chromatography can be used to distinguish between different biomolecules based on their chemical
10 Pages(2500 words)Lab Report
Biochemistry lab report
The ratio of the distance covered by the molecule to be separated and by the solvent is called the relative front and is used to identify an unknown amino acid. Since the amino acids
10 Pages(2500 words)Lab Report
Lab report
Then place another clamp with a 200-g suspension and repeat the same previous step. Record positions with respect to the points of support and of the weights used. Empty the meter stick of clamps and
3 Pages(750 words)Lab Report
Lab Report
1. E. coli is in competition with S. euglensis at a temperature of 20-35 degrees and a pH of 6-7.5. E. coli is also in competition with N. atol at a temperature of 20-30 degrees and a pH of 4.5 -6.5. P.
1 Pages(250 words)Lab Report
Enzymes lab report
The results show that lactase activity is low when temperature is low at 00C but improves with increasing temperature up to 370C after which activity drops sharply. Low activity at low temperature is due to low activation energy exhibited by
5 Pages(1250 words)Lab Report
Lab report
For the first cycle, the mass of the water alone is the mass of the crucible lid and hydrate, less the mass of the crucible lid and residue after heating. That gives 22.678 – 22.460 = 0.218. The mass of the hydrated salt is the mass of the crucible lid and hydrated salt, less
1 Pages(250 words)Lab Report
Lab report
o calibrate the pressure transducer and then perform an uncertainty analysis and compare your pressure transducer calibration to the manufacturer’s calibration. 1. Excite the pressure transducer, with a voltage of 10v. The figure above had J17 and J18 as AO (Analog Out). It
5 Pages(1250 words)Lab Report
Lab formal report
The law is bellow, The primary aim of the underlying experiment is to determine the molecular mass of a volatile liquid in weighed flask. A small amount of the
1 Pages(250 words)Lab Report
Lab report
e results of the experiment indicated that biuret protein analysis methods is a more definitive method of protein determination as the concentration mean of protein using this method was 1.88 x 10-3 while Folin protein assay methods had a standard deviation of 0.026. Biuret
4 Pages(1000 words)Lab Report
Let us find you another Lab Report on topic Biochemistry Lab Report for FREE!
Contact us:
Contact Us Now
FREE Mobile Apps:
  • About StudentShare
  • Testimonials
  • FAQ
  • Blog
  • Free Essays
  • New Essays
  • Essays
  • The Newest Essay Topics
  • Index samples by all dates
Join us:
Contact Us