Restriction Enzyme Analysis
Research question: Using only the information from these experiments
calculate the number and size of the fragments that would be made
using BAMH1 restriction enzyme, and calculate the migration distances
of the various fragments.
Restriction enzyme used
Base pairing fragments
Distance Travelled (cm)*
Log of base pairs*
Example
My Group
ECO R1 (6 fragments)
21226
7421
5804
5643
4878
3530
1.4
2.0
2.3
2.5
2.9
-
1.7
2.0
2.2
2.3
-
-
4.3
3.9
3.8
3.8
3.7
3.5
HIND III (7 fragments)
23130
9416
6682
4361
2322
2027
564
1.4
1.8
2.2
2.7
3.5
3.7
5.8
1.6
1.8
2.1
2.3
2.9
3.1
-
4.4
4.0
3.8
3.6
3.4
3.3
2.8
Predicted figures from my graph
BAM H1 (6 fragments)
16842
7233
6770
6527
5626
5504
1.3
2.2
2.5
2.5
2.5
2.8
4.2
3.9
3.8
3.8
3.8
3.7
Results
*All figures in table are to 2s.f.
I chose to calculate the log of the base pairs because the numbers
vary significantly, making it difficult to plot on a graph. By doing
this you can put the values into perspective for easy comparison. I
will only use the example on my graph, as my results are unclear and
inaccurate. I measured the distance from the edge of the gel, where
the well ended to the centre of each fragment.
The main purpose of this experiment is to examine the results of wild-type mutant crosses which influence the arrangements of ascospores in asci in the fungus Sordaria fimicola. These resulting arrangements help calculate the map distance between the centromere and spore color genes in Sordaria. My hypothesis was that due to so many group observations accounted in, the data will be underestimated and the results will not fit into the chi square table. A sample from Petri dish with both mutant stock cultures is observed after a week. The ascospores must appear in MII pattern 2:2:2:2 or 2:4:2 arrangements in order for the crossing-over to occur. Next, based on the data collected, the class calculated the map distance. If the map distance does not fit the value obtained by the researchers from the many successful experiment attempted, then the experiment had errors. And due to this, the class experiment cannot accept the null hypothesis according to the chi square test. However, our class experiment accepted the null hypothesis and so it was a success.
place 3cm depth of medium sucrose solution (0.5 Molar) into a boiling tube and label C 4. place 3cm depth of stronger sucrose solution (0.75 molar) into a boiling tube an label D 5. place 3cm depth of strong sucrose solution (1 Molar) into a boiling tube and label E Here is a table of how to make the sucrose solution: concentration sucrose Water 0M 0ml 10ml 0.25M 2.5ml 7.5ml 0.5M 5ml 5ml 0.75M 7.5ml 2.5ml 1M 10ml 0ml Then I will: 1. Cut 5 potato chips; accurately measure the length (mm) and mass (g) and record these results 2. place one chip into each tube and leave for 20mins 3.
Use the ruler to measure the distance of the opisthocranion orale in cm,then record your measurements
As the gelatin cube sizes were cut into smaller pieces, the surface area to volume ratio increases. Surface area to volume ration is very important in living organisms, since all the nutrients and oxygen need to diffuse through the cell membrane into the cells. The surface area to volume ratio affects the rate in which the salt diffused through the solution by how the salt diffused out of the gelatin and into the water this relates to small cells that have a faster rate of diffusion for the nutrients and oxygen to diffuse into the cell and out of the cell quicker. Whereas if the cells were bigger, the rate of diffusion for the nutrients and oxygen to diffuse into the cell would take too long and the cell would probably not survive. So the surface area to volume ratio in cells is vital since the smaller they are the faster the rate of diffusion of the nutrients and oxygen can go in and out for the cell to be able to grow and survive.
The replicated DNA is then treated with restriction enzymes, which cut the DNA at their specific sites. Purpose The purpose of this lab was to understand how to analyze a series of genetic data from DNA using agarose gel electrophoresis and decode the results. Materials Micropipette Micropipette tips Premade Agarose gel Gel electrophoresis machine DNA samples ‘Marker’ sample UV light box Methods 1.
You can make sure to keep the size of the magnesium ribbon constant throughout the experiment by measuring it accurately with the help of a ruler. Make sure that the length of the magnesium ribbon is 2 cm throughout the experiment.
The effects of temperature in enzymes ABSTRACT This report explains enzymes and how their activity can change depending on different factors. We explored this by breaking the experiment into two parts; in the first half we measured peroxidase activity at the different given temperatures and the second half we repeated this but let it sit at room temperature before measuring in the spectrophotometer again. The results showed that temperature does affect the ability of peroxidase. The effects were not reversible.
The mixture for that table’s flask was 15 mL Sucrose, 10 mL of RO water and 10 mL of Yeast, which the flask was then placed in an incubator at 37 degrees Celsius. In my hypothesis for comparison #4 the measurements would go up again with every 15 min. intervals because of the high tempeture and also be higher that then Controlled Table’s measurements. Hypothesis was right for the first part but was wrong for the second part of the comparison, the measurements did increase in the table’s personal flask but the measurements did not get higher than the Controlled Table’s measurements, see chart below. In conclusion, I feel that the substitution of glucose for sucrose made the enzymes work just as hard as the Controlled Table’s flask but just not as much because sucrose was too strong for the enzymes to
The procedure is very short. First, I constructed the data table. I then massed one piece of Aluminum foil and record it in the data table. Next, pour 25.0 mL of 0.400 M Cu²+ solution and tear up the massed piece of foil. Drop the torn pieces in the solution and record the observations. Mass the other pieces of Aluminum foil, tear it up, and drop it in 25.0 mL of the 0.200 M Cu²+ solution. Record observations and clean up according to the teachers
My hypothesis was I think that if we change the temperature of the enzyme it will have an area where the enzyme works very well, but if we increase or decrease the temperature too much, it will slow the productivity. If we change the pH of the enzyme, there will be an area where it is extremely productive, but if it gets too acidic or basic the productivity will slow. If we change the concentration by increasing the amount of enzymes, it will be more productive. T think this because your are having more than 1 enzyme work on one task. I you decrease the amount of enzymes, the productivity will be slower because there are less enzymes working on that certain task. The data refutes my hypothesis because I found that the enzymes work best in one
3. Place the end of the ruler at one edge of the crater and measure to
The discovery and characterization of restriction enzymes first took place in the late 1960’s and early 1970’s. The scientists responsible for the discovery were molecular biologists Werner Arber, Hamilton Smith and Daniel Nathans. In the late 60’s Arber observed a sharp change in the bacteriophage DNA he had been working with after it invaded resistant strains of bacteria. It had been cut into pieces and degraded. He hypothesized that bacteria could express two different enzymes: one that recognizes and destroys foreign DNA, the restriction enzyme, and one that modifies bacterial DNA to protect it from the former, a modification enzyme. A short time later he, along with Stuart Linn, confirmed his second hypothesis that both enzymes act on the same specific sequence of DNA, the recognition sequence. In 1970 Hamilton Smith both verified and elaborated on Arber and Linn’s hypothesis and initial discovery using a
One of the best characteristics for the functional status of a certain cell is its gene expression pattern. Cells belonging to different tissues, cells in different developmental or metabolic stages, cells under the influence of specific compounds, or cells within a carcinogenic process differ by their gene expression patterns and thus by their mRNA pools. Currently, the most important technique for the accurate quantitation of gene expression is the fluorescent quantitative real-time RT-PCR (Muller et al., 2002a).
An alternative approach of affinity chromatography with extremely significant results is dye-ligand affinity chromatography. In this type of affinity chromatography, dyes compose the group of ligands than are employed in the technique (Hage et al., 2012).
Begin collecting samples with the pure hexane. Keep adding hexane so that the silica gel column does not run dry. Collect one 20 ml sample. Repeat with 90:10 hexane and collect 4 20-mL bottles. Repeat with 80:20 hexane and collect 2 20-mL samples.