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Any SINE of Chromosome 16? PV92 PCR/Informatics ( S hort In terspersed Repetitive E lement)

Any SINE of Chromosome 16? PV92 PCR/Informatics ( S hort In terspersed Repetitive E lement). Think of your entire genome as one very fat book of biological data. Like this. 100 pages total: - 95 pages of nonsense - 5 pages of useful information.

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Any SINE of Chromosome 16? PV92 PCR/Informatics ( S hort In terspersed Repetitive E lement)

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  1. Any SINE of Chromosome 16? PV92 PCR/Informatics (Short Interspersed Repetitive Element)

  2. Think of your entire genomeas one very fatbook of biological data. Like this...

  3. 100 pages total: - 95 pages of nonsense - 5 pages of useful information

  4. Only 5 pages of your genetic book contain useful information!

  5. Today we are going to look in a non-coding region of your chromosome #16 But remember: You got one chromosome #16 from Mom And one chromosome #16 from Dad Mom thinks hers is better.

  6. Alu repeats: our targets • Classified as SINEs (Short Interspersed Repetitive Element) • Mobilized by an RNA polymerase-derived intermediate (retroposition) • Approx. 500,000 Alu copies per haploid genome, representing about 5% of the genome • Named for the Alu I restriction site within the element

  7. Evolutionary Significance of PV92 Alu Inserts Highly conserved Inserted in the last 1,000,000 years Genotypes (+/+, +/-, -/-) Used in population genetics, paternity analysis, and forensics

  8. Alu 3’ 5’ Amplified Region PCR Results • The PV92 Alu is dimorphic so there are two possible PCR products: • 641 bp • 941 bp No insertion: 641 bp Alu insertion: 941 bp 641 bp 300 bp Alu insert

  9. Actual Alu PCR Results - +/- + 941 bp 641 bp + - +/-

  10. Amplify Alu insert from representative sample population Calculate the expected allelic and genotypic frequencies To estimate frequency of Alu within a population:

  11. InstaGene Extraction Cell membrane Nuclear membrane Mg++ Genomic DNA Mg++ Mg++ Mg++ Heat disrupts membranes Mg++ InstaGene matrix binds released cellular Mg++ Mg++

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