Know thy self Luxury Genes - from Forbes.com Gift-hunting for a truly self-obsessed friend? Consider buying him/her a reading of his/her entire DNA code.

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Presentation transcript:

Know thy self Luxury Genes - from Forbes.com Gift-hunting for a truly self-obsessed friend? Consider buying him/her a reading of his/her entire DNA code A firm in Cambridge, Mass. called Knome (pronounced "gnome") is offering the service. Recent News October 2008: Knome Commences Whole-Genome Sequencing Process For First Clients Commences Whole-Genome Sequencing Process For First Clients

You, on a platter: A customer who pays $350,000 for Knome’s service receives a silver box containing an eight-gigabyte USB drive housing his or her genome sequence and specialized browser software. Credit: courtesy of Knome, Inc.

PCR Amplifies Target DNA in vitro Replicating target DNA in vitro Taq polymerase Temperature-resistant DNA polymerase –Thermus aquaticus Heat resistant Best for <2 kb target 2581b Hill C9

Fig PCR Replicating target DNA in vitro 2581b Hill C9

Fig Two primers bracket the target region to be amplified Complementary to one strand Anti-parallel Amplify DNA PCR 2581b Hill C9

Fig Amplify DNA PCR The 3’ nucleotide is critical for specificity 2581b Hill C9

Fig Amplify DNA PCR Cycles of repeated replication 2581b Hill C9

Fig Denature Anneal Extend 2581b Hill C9

Primer Artefact Lane M N P kb What is the size of the PCR product? Gel electrophoresis separates PCR amplicons on the basis of size. 2581b Hill C9

Exponential increase Need to know some sequence information Very sensitive Can amplify from limited starting material Ideal for 2kb target PCR Fig b Hill C9 The PCR song Lads&feature=related

Fig DNA sequencing 2581b Hill C9

Fig DNA sequencing 2581b Hill C9

Fig DNA sequencing ddTTP 2581b Hill C9

Original Strand / Template strand Nascent or New Strand Autoradiograph 2581b Hill C9

Fig b Hill C9

Fig a KH Genomics 10

Fig d Automated DNA Sequencing KH Genomics 10 Gels are now replaced by capillaries filled with polymers

Sequencing Chromatogram KH Genomics 10 5’ 3’ Nascent or Newly synthesized strand Close to the sequencing primer Far from the sequencing primer

Page 349 Heterozygote Detection Maternal template sequence A Paternal template sequence C SNP Single nucleotide polymorphism KH Genomics 10

Page 349 Heterozygote Detection KH Genomics 10 What would a chromatogram look like if the maternal copy had a deletion of one nucleotide? A typical DNA sequencing read is 1000 nucleotides