Preview

Rlfp

Powerful Essays
Open Document
Open Document
1429 Words
Grammar
Grammar
Plagiarism
Plagiarism
Writing
Writing
Score
Score
Rlfp
Restriction Fragment Length Polymorphism (RFLP)
Restriction Fragment Length Polymorphism (RFLP)

Definition
Definition

In molecular biology, restriction fragment length polymorphism, or RFLP is a technique that exploits variations in homologous DNA sequences. It refers to a difference between samples of homologous DNA molecules that come from differing locations of restriction enzyme sites, and to a related laboratory technique by which these segments can be illustrated. In RFLP analysis, the DNA sample is broken into pieces (digested) by restriction enzymes and the resulting restriction fragments are separated according to their lengths by gel electrophoresis. Although now largely obsolete due to the rise of inexpensive DNA sequencing technologies, RFLP analysis was the first DNA profiling technique inexpensive enough to see widespread application. In addition to genetic fingerprinting, RFLP was an important tool in genome mapping, localization of genes for genetic disorders, determination of risk for disease, and paternity testing. Restriction Fragment Length Polymorphism (RFLP) is a difference in homologous DNA sequences that can be detected by the presence of fragments of different lengths after digestion of the DNA samples in question with specific restriction endonucleases. RFLP, as a molecular marker, is specific to a single clone/restriction enzyme combination. Most RFLP markers are co-dominant (both alleles in heterozygous sample will be detected) and highly locus-specific. An RFLP probe is a labeled DNA sequence that hybridizes with one or more fragments of the digested DNA sample after they were separated by gel electrophoresis, thus revealing a unique blotting pattern characteristic to a specific genotype at a specific locus. Short, single- or low-copy genomic DNA or cDNA clones are typically used as RFLP probes.The RFLP probes are frequently used in genome mapping and in variation analysis (genotyping, forensics, paternity tests, hereditary

You May Also Find These Documents Helpful

  • Good Essays

    This was done by melting a solution composed of 0.7% agarose in 1x TBE and 0.005% ethidium bromide, and pouring the resulting liquid into comb-containing gel tray until the thickness reached about 6 mm. Once the gel had been allowed to cool for about 20 minutes, it was removed from the tray, placed in a horizontal gel apparatus, and immersed in 1x TBE buffer. Next, three different solutions were prepared from an aliquot of HindII digested lambda DNA and 3 microliters of 6x loading dye, an aliquot of EcoRI digested lambda DNA and 3 microliters of 6x loading dye, and 1 kb DNA ladder and 3 microliters of 6x loading dye. The resulting solutions were then transferred into separate wells within the gel, and the apparatus was connected to a constant current source (45mA) for 60 minutes. After producing clearly visible bands, the gel was taken out of the apparatus and photographed with an ultraviolet light box (Displayed in Figure…

    • 1568 Words
    • 7 Pages
    Good Essays
  • Better Essays

    Apush Unit 5 Review

    • 1238 Words
    • 5 Pages

    The purpose of this lab was to observe the different amount of distances each DNA samples travel when placed in a gel-electrophoresis box. Restriction endonucleases are critical tools in recombinant DNA methodology. Electrophoresis is the method of determining the size of fragments that are cut by restriction enzymes. These restriction enzymes always cut at their specific protein recognition sites. This is very useful in the sense that no two restriction enzymes codes for exactly the same recognition site, giving it a unique characteristic that is specific for a strand of DNA. Gel electrophoresis is a technique used to separate different sized fragments of DNA or RNA with the use of an electric field. When a molecule enters an electric field, the speed at which the molecule moves is influenced by several factors including: the charge of the molecule, the strength of the electrical field, the size and shape of the molecule, and the density of the medium (in this case the agarose gel) through which the molecule moves. Because of this, scientists are able to separate different groups of DNA or RNA molecules by first positioning all the molecules at a uniform starting point on the agarose gel and then placing the gel in a chamber containing buffer solution and electrodes. A buffer is a solution that adds extra ions to the gel enhancing the conductivity in the agarose gel matrix. Once in the chamber containing buffer solution and electrodes, the molecules of DNA will begin to migrate through the gel and form bands due to the negative charge of the phosphate groups in the backbone of DNA moving towards the positive electrode.…

    • 1238 Words
    • 5 Pages
    Better Essays
  • Good Essays

    Through the process of gel electrophoresis, DNA fragments are able to be separated. Gel electrophoresis is a method of separating and analysis DNA molecule fragments based on their size and charge. On end of the gel is given a positively charged end and one end is negatively charged. When an electric current is passed through the gel charged molecules move through it. Larger molecules move slower, moving a shorter distance, while smaller molecules move faster and traveler further. These DNA molecules are separated by size in the gel and dye is used to stain the fragments and make them more visible.…

    • 231 Words
    • 1 Page
    Good Essays
  • Good Essays

    of genomic DNA that has been cut with restriction enzyme X. The gene you wish to insert has…

    • 4889 Words
    • 20 Pages
    Good Essays
  • Powerful Essays

    In PBS, you learned about the molecular biology techniques that allow scientists to explore our DNA. PCR, Polymerase Chain Reaction, is the copy machine; the revolutionary process that allows scientists to replicate even the tiniest speck of DNA. Restriction endonucleases (enzymes) are the molecular scissors that can cut DNA in specific locations. Your specific code determines the number of times this set of scissors will snip and the number and size of DNA pieces that will be left behind. These pieces can then be separated and compared using the process of gel electrophoresis. As these fragments move, their varying lengths propel them through the gel at different speeds. Scientists can use these RFLPs, Restriction Fragment Length Polymorphisms, a set of DNA puzzle pieces unique to only you, to create a pattern called a DNA fingerprint. Similar to the unique…

    • 1747 Words
    • 6 Pages
    Powerful Essays
  • Good Essays

    An individual’s code determines the number of times the restriction enzymes will cut and the number and size of DNA pieces that will result. These pieces can then be separated and compared using a process called gel electrophoresis. The DNA moves from the negative end to the positive end. As the fragments move, their varying lengths propel them through an agarose gel at different speeds. Short strands move through the holes in the gel more quickly than long strands and will over time move farther away from the starting point. Staining the sorted groups of DNA makes them visible to the naked eye (show up as bands in the…

    • 1489 Words
    • 6 Pages
    Good Essays
  • Good Essays

    Lab 8: Genetic Analysis

    • 297 Words
    • 2 Pages

    In Lab 8, we had analyzed remains found at a wooded area near Jonesburg and tried to determine if the bones belonged to a 28-year-old woman who had been reported missing from a city within the vicinity. Upon analysis, it was determined that they did belong to a female. However, it was not possible to determine if the bones did belong to the missing women. Lab 12 presented the opportunity to genetically analyze the remains found. DNA profiling, also referred to as typing and fingerprinting, uses genetic material to show relatedness and uncover the identity of organisms. Most commonly associated with forensics, it can be used in an array of scientific fields such as anthropology. One method that can be used, when a large sample present, is restriction…

    • 297 Words
    • 2 Pages
    Good Essays
  • Satisfactory Essays

    Forensic 1 Unit 7 Text

    • 260 Words
    • 1 Page

    5. What is RFLP? What are some of the limitations of this technique? RFLP is a method used by molecular biologists in order to follow a sequence enzyme.…

    • 260 Words
    • 1 Page
    Satisfactory Essays
  • Good Essays

    Restriction enzymes are tools in DNA research that can cut DNA into exactly needed pieces. Certain cuts can be rough, while others can be clean. Certain cuts can have an organized pattern to have a staggered cut. Other cuts will leave complementary bases with them. Electrophoresis allows the manipulation of DNA to separate and organize those parts. Electrophoresis is the substrate electric movement of the separation of DNA. Gel Electrophoresis can separate both DNA and RNA. This can also allow the knowledge of the unknown bases of a strand of DNA. This can allow the knowledge of familiar patterns that can create a unique signature. Separation of these molecules…

    • 1180 Words
    • 5 Pages
    Good Essays
  • Good Essays

    A sample of DNA found in a crime scene was provided along with five suspects. Their DNA was then processed using restriction enzymes and Agarose Gel Electrophoresis. The objective of this lab was to match a criminals DNA to a crime scene using restriction enzymes EcoRI and Pstl with Agarose gel electrophoresis. Restriction enzymes cut DNA at a specific base pair site recognized by the enzyme, which then turns one single strand of DNA into many fragmented strands of DNA. EcoRI recognizes and cuts the palindromic base pair sequence GATTC while Pstl recognizes and cuts the palindromic base pair sequence CTGCAG. Agarose gel electrophoreses separates these fragmented DNA by their size. The negatively charged DNA moves through the Agarose gel to the positively charged end of the gel. The smaller fragments move through the gel more quickly allowing a linear view of the fragmented DNA when the process is complete. Since each individuals DNA will be cut into different size fragments when restriction enzymes are applied we can match one of the suspects to the crime scene DNA sample. This process enables an individual’s DNA to be matched, much like a fingerprint, to a sample of unknown DNA.…

    • 866 Words
    • 4 Pages
    Good Essays
  • Better Essays

    DNA Fingerprinting

    • 1281 Words
    • 5 Pages

    After the restriction digest incubation period, are there any visible clues that the restriction enzymes may have in some way changed the DNA in any of the tubes Explain your reasoning. Day 2 Post Lab Observation of Results Sketch your result to the left. Dry your gel as instructed. Quantitative Analysis of DNA Fragment Sizes If you were on trial or were trying to identify an endangered species, would you want to rely on a technicians eyeball estimate of a match, or would you want some more accurate measurement In order to make the…

    • 1281 Words
    • 5 Pages
    Better Essays
  • Powerful Essays

    In biology restriction enzymes are used in several ways to modify and manipulate DNA molecules. One common use is to compare pieces of DNA from one that is unknown, with fragments of DNA from another source that the lengths are known. This process requires preparation of small fragments that are appropriate for nucleotide sequence analysis is to provide a map of the distribution of target sites. The cuts occur when palindromes show up in the DNA sequence; the different restriction endonucleases cut these different sites. This is called constructing a restriction map. In this experiment we are given an unknown DNA and use the process of DNA mapping with gel electrophoresis to compare and identify if the given DNA plasmid is of this planet and to describe what the DNA map illustrates.…

    • 1175 Words
    • 5 Pages
    Powerful Essays
  • Satisfactory Essays

    Cafs Irp

    • 827 Words
    • 4 Pages

    Create an Independent Research Project by conducting your own research on a particular subject/focus and construct a report detailing your research.…

    • 827 Words
    • 4 Pages
    Satisfactory Essays
  • Satisfactory Essays

    Cafs Irp

    • 987 Words
    • 4 Pages

    I chose this topic because it is relevant to the Community and Family Studies course and because I am extremely interested in this focus area. This chosen research question is relevant to the Community and Family Studies course. These of which include:…

    • 987 Words
    • 4 Pages
    Satisfactory Essays
  • Good Essays

    The DNA profiling and its interpretation have come under fire. RFLP analysis was in part discontinued because of the possibility for error. The risk of a coincidental match using RFLP is 1 in 100 billion. However, in laboratory settings, this risk is probably higher because technicians may misread…

    • 576 Words
    • 2 Pages
    Good Essays

Related Topics