ipl-logo

Restriction Enzyme Lab Report

866 Words4 Pages

Function of Restriction Enzymes:
Restriction endonucleases cleave the phosphodiester bond between an adjacent phosphate and deoxyribose group in the phosphate backbone of the DNA. The active site of the endonuclease perform this cleavage by binding to the side chain of certain amino acids to the phosphate group through a chemical bond. This dissolves the preexisting bond between the deoxyribose sugar and the phosphate resulting in a breakage with in the DNA chain at a specific location. (3, 7)
One characteristic feature of restriction endonucleases is that they cut at a very particular site having a specific DNA sequence. This specific sequence that allows the enzyme to attach is known as the recognition site. Consider the example of the first restriction enzyme discovered, EcoRI. …show more content…

Pulse field electrophoresis is used for the separation of larger fragments of DNA. These fragments result from digesting a bacterial genome with a rare-cutting restriction enzyme. The pattern of DNA produced on the gel is used to differentiate different strains of bacteria. For the identification of individuals like humans or other organisms, Restriction fragment length polymorphism (RELP) analysis has turn out to be very practical. After isolation of DNA from the source it is digested enzymatically with the help of restriction endonucleases. Enzyme treated DNA is then separated by size in an agarose gel and shifted to a membrane. A radioactive or fluorescently labeled probe is bonded with the DNA on the membrane. They target specific sequences that are marked by the restriction enzyme sites. The size of these fragments varies hence generate a biological bar code of restriction enzyme- digested DNA fragments. This pattern is unique to each individual. Restriction enzymes are fore sighted to be an integral part of the modern genetics. (3,

Show More
Related
506 Words | 3 Pages

Tn 4351 was originally isolated from bacteroides fragilis [30] . The transposon was successfully introduced into Cytophaga succinicans, Flavobacterium meningosepticum, Flexibacter canadiansis, Flexibacter strain SFI and Sporocytophaga myxococcoides by conjugation [25]. Tn 4351carries two antibiotic resistance gene. One of the codes for resistance to erythromycin and clindamycin which is expressed in bactroides but not in E.Coli. The other gene codes for resistance in tetracycline and is expressed in aerobically grpwn E. coli, but not in anaerobically grpwn E. coli or in bacteroides.

Read More
735 Words | 3 Pages

How could a single nucleotide change on the plasmid alter the fragment pattern observed in the gel? (3 points) When a single nucleotide is change on the on the plasmid, it will make the restriction enzyme not to recognize the fragment pattern and also all the numbers at each site will be altered or this action could make the fragment not to be incised at the expected site, which could make a larger fragment to travels a short interval on the gel with less fragments than expected. It could also make it to be incised at more places than that originally expected and the result will be increased, smaller fragments that move further across the gel. There may also be no change in the banding pattern.

Read More
944 Words | 4 Pages

After multiple cycles of ligation, detection and tail cleavages, the extended chain reached the end of the template. Then the whole extension chain is removed and a new starting primer switching down 1 nucleotide position binds onto the template for another cycle of reaction. Totally, five round of primer binding cycles are performed to complete the sequencing of each fragment. 3. Pitfalls and limitations of NGS Errors could be introduced in any step of the sequencing process, including library

Read More
494 Words | 2 Pages

The competitive inhibitor that was added was lactose. We predicted this because competitive inhibitors block and bind to the active site so it will slow down the binding of the desired substrate. An alternative hypothesis that came up was that the reaction of substrate would stay consistent as if no inhibitor was added. The enzyme could reject the inhibitor if it does not fit in the active site, causing the substrate to bind as it normally would. Our results showed that with the addition of lactose, the reaction did slow down a considerably

Read More
763 Words | 4 Pages

Once the loading solutions were made and the agarose gel electrophoresis apparatus was set up, the solutions were dispensed into the wells of the gel, as well as a reference solution. The gel image gathered from the gel electrophoresis allowed for the analysis of the plasmid DNA and the effects the restriction endonucleases, Hind III and Hinc II, had on it. While analyzing the gel image, it was apparent that based on the reference solution in lane 1 and the mark in lane 3, the Hind III solution, that the number of base pairs should be the same. Also it was apparent that lane 5, the Hinc II solution, had 4 marks. By measuring the migration distance in centimeters and plotting against the log(# of base pairs(bp)) for the standard in lane 1 the linear trendline equation provided by the calibration curve allows for the number of base pairs to be found for the marks in lane 3 (Hind III) and lane 5 (Hinc II).

Read More
503 Words | 3 Pages

Investigating DNA Aim: To extract and examine DNA. Materials: As per page 40. Method: As per page 40.

Read More
1168 Words | 5 Pages

The STR length contrast is what is used to differentiate individuals. Gel electrophoresis then uses the STRs to create a DNA profile. The gel electrophoresis separates the STRs depending on their length and the pattern is then shown in fluorescent gel creating the profile. These profiles are then used by scientist to compare patterns between evidence and or suspects to determine a match or not a match.

Read More
450 Words | 2 Pages

When scientist began studying the way that bacterial protect themselves from different types of viral and from bacteriophages, they realize that inside bacterial cells are these special digesting proteins and special digestive enzymes known restriction enzymes, these restriction enzyme are able to a cut or cleave the viral DNA molecule into many different pieces thereby destroying the viral DNA and deactivating the viral DNA. There are many different types of restriction enzymes that exist in nature and each restriction enzyme cleaves a longer DNA molecule at a specific location on that double stranded DNA molecule. Different types of restriction enzyme also recognize a different sequences in a DNA and and it will cut in different way. Restriction enzyme cuts the DNA

Read More
299 Words | 2 Pages

The portion of the genome that codes for a protein or an RNA is referred to as

Read More
614 Words | 3 Pages

This leads to the production of a corepressor, which eventually binds to the repressor and activates it. Inducible operons, like the lac operon, are normally off with active repressors. However in the presence of lactose, the inducer allolactose binds to the repressor, making it inactive and turning the operon on. This leads to the transcription of the genes that code for proteins β-galactosidase, permease, and transacetylase. b. Four mechanisms of eukaryotic gene regulation are the 5’ cap, which prevents degradation, the poly A tail, which allows exiting from the nucleus, the alternate splicing of introns by spliceosomes, and acetylation of histones, which makes DNA less condensed.

Read More
589 Words | 3 Pages

This method uses RFLPs as they travel across a gel, attracted to the other side of the gel with a positive charge. What are restriction enzymes? A restriction enzyme is an enzyme that cuts DNA at only certain (but can be palindromic) sequences in the DNA.

Read More
1294 Words | 6 Pages

The improvement of DNA sequencing instruments, detection mechanisms and bioinformatics all contribute to the development of automated DNA sequencing. The human genome sequencing projects provided a

Read More
1789 Words | 8 Pages

Single digestion is only one restriction enzyme which has been used to digest a DNA. Double digestion is there are 2 restriction enzymes which have been used to digest a DNA. Based on the result, in the gel electrophoresis, P1-1 has form two bands as it has 2 restriction sites.

Read More
657 Words | 3 Pages

Two of them are direct DNA mutation analysis, and indirect DNA testing. In direct DNA mutation analysis, blood is usually drawn from a person. This test is only possible if a gene mutation is known. Chemicals are used to separate the DNA from the cell. Heat separates the strands, and enzymes called restriction enzymes are placed to cut the strands in specific places.

Read More
1251 Words | 6 Pages

Finally, the amplified DNA regions are compare using a gel. DNA Profiling

Read More

More about Restriction Enzyme Lab Report

Related Topics

Open Document