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    MT DNA AND SEQUENCING

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    Nived K K
    Mitochondria contain circular DNA molecules that are passed down solely from mothers. Mitochondrial DNA profiling can be used to determine maternal relationships by sequencing a non-coding region called the D-loop. This technique is useful for identity testing from old, degraded, or limited samples when nuclear DNA cannot be obtained. The sequencing process involves extracting, quantifying, amplifying via PCR, sequencing the amplified DNA, and comparing the results to reference sequences.

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    MT DNA AND SEQUENCING

    Uploaded by

    Nived K K
    15 views2 pages
    Mitochondria contain circular DNA molecules that are passed down solely from mothers. Mitochondrial DNA profiling can be used to determine maternal relationships by sequencing a non-coding region called the D-loop. This technique is useful for identity testing from old, degraded, or limited samples when nuclear DNA cannot be obtained. The sequencing process involves extracting, quantifying, amplifying via PCR, sequencing the amplified DNA, and comparing the results to reference sequences.

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    Mtdna

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    Mt DNA AND SEQUENCING

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    Mitochondria contain circular DNA molecules that are passed down solely from mothers. Mitochondrial DNA profiling can be used to determine maternal relationships by sequencing a non-coding region called the D-loop. This technique is useful for identity testing from old, degraded, or limited samples when nuclear DNA cannot be obtained. The sequencing process involves extracting, quantifying, amplifying via PCR, sequencing the amplified DNA, and comparing the results to reference sequences.

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    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    15 views2 pages

    MT DNA AND SEQUENCING

    Uploaded by

    Nived K K
    Mitochondria contain circular DNA molecules that are passed down solely from mothers. Mitochondrial DNA profiling can be used to determine maternal relationships by sequencing a non-coding region called the D-loop. This technique is useful for identity testing from old, degraded, or limited samples when nuclear DNA cannot be obtained. The sequencing process involves extracting, quantifying, amplifying via PCR, sequencing the amplified DNA, and comparing the results to reference sequences.

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    © All Rights Reserved
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    Mitochondrial DNA

    ● Mitochondria are body gene contained in eukaryotic cells, responsible for cellular
    respiration and for the generation of Adenosine triphosphate (ATP)-an important body
    constituent for the transfer of energy in living cells.
    ● Mitochondria genes give mitochondrial DNA (mtDNA).
    ● The mitochondrial genes are inherited from mother only.
    ● Mitochondrial genes cannot be used to determine the fatherhood of a person or child.
    ● They, however, link mother and other maternal relatives positively.

    CHARACTERISTICS
    ● Human mitochondria contain 2 to 10 units of double stranded circular DNA molecule.
    ● The mitochondrial genome is highly variable.
    ● Human mtDNA contains16569 base pairs.
    ● These code for 37 genes that in turn code for the synthesisof 2 ribosomal RNAs, 22
    transfer RNAs and 13 proteins.
    ● Unlike nuclear DNA,the mitochondrial genome is extremely compact and about 93 per
    cent of theDNA represents coding sequences. The remaining, non-coding region is
    called the control region or displacement loop (D-loop). The D-loop region consists
    ● D-loop. provides high discrimination among non-related individuals on analysis with
    restricted enzymes.
    ● It has also been found that maternally related individuals show abundant
    correspondence in D-loop sequences.
    ● The D-loop segment is amendable to polymerase chain reaction and, thus, minute
    quantities of the mtDNA can provide mitochondrial DNA profiling for determining
    maternal interrelationship or identity.

    SIGNIFICANCE
    MtDNA profiling is coming up as an important tool to establish the identity or non-identity of an
    individual from hair (shaft), teeth and bones even when they are old. Mr DNA has thus widened
    the identification field tremendously.
    It is very useful when
    ● Nuclear DNA is significantly degraded.
    ● Nuclear DNA is present in small quantities.
    ● Authorities cannot obtain a reference sample from an individual who is looking deseased
    or missing.

    Mt DNA SEQUENCING
    The sequencing and comparison of mtDNA can be broken up into five steps.
    1. Extraction
    2. Quantitation
    3. PCR
    4. Sequencing
    5. Comparison
    EXTRACTION
    QUANTITATION
    PCR ANALYSIS
    Using this technique many copies can be produced from a relatively small number of copies.
    SEQUENCING
    Gel Electrophoresis:
    ● DNA products are separated by length of bp
    ● pore size of the gel influence how far the DNA fragments will travel when placed in an
    electric field
    ● smaller fragments will travel faster and appear further from the wells in the gel
    ● larger fragments will travel slower and appear closer to the wells in the gel
    ● fluorescent detector "records the emitted wavelength of the fluorescent dyes on each
    base as the fragments travel past the detection area of the instrument"
    ● a chromatogram is generated, showing the colors of the labeled fragments
    ● the sequence of the mtDNA is determined from series of cycle sequencing a reactions

    COMPARISION
    ● mtDNA sequences are generated by a computer and edited by a DNA examiner to
    obtain the final sequence
    ● Difference(s) is/are recorded by comparing the finalized sequence to the reference
    sequence
    ● If sequence concordance ("the presence of the same base or a common base at every
    position analyzed") is observed, then both mtDNA samples could be considered as
    originating from the same source

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