Bio 142 - Week 3 slides - Sp12-4

Bio 142 - Week 3 slides - Sp12-4 - PCR and Bioinformatics:...

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PCR and Bioinformatics: Tools you can use!
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PCR Experiment: VNTR Tube Add 22.5 µL of pMCT118 primer/loading dye mix (dyed pink) Tap/mix until bead dissolves Label the cap and side of your tube with the number assigned by your instructor. Spin down any solution that is on the side of your tube and store on ice until all your tubes are prepared. Add 2.5 µL of YOUR DNA and mix L. Mclane and A. Escobar
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RFLP of Exon 6 and 7 PCR products L. Mclane and A. Escobar
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L. Mclane and A. Escobar BLOOD TYPE PCR
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L. Mclane and A. Escobar Restriction Enzyme Recognition Sites: Nar I: 5’…GG*CGCC…3’ 3’…CCGC*GG…5’ Pml I: 5’…CAC*GTG…3’ 3’…GTG*CAG…5’ Kpn I: 5’…GGTAC*C…3’ 3’…C*CATGG…5’ *Indicates Enzyme Cut Site The patterns of the bands that result from restriction digests in exons 6 and 7 will determine blood type. The digests will produce fragments of different sizes; the presence of these different bands will determine your blood type! RFLP Analysis of Blood Type
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RFLP Analysis of Individual with Type A Blood PCR-RFLP successfully genotyped this individual as AO 1 . Wheeler and Waikel
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L. Mclane and A. Escobar VNTR - D1S58 - defined by DNA probe pMCT118 VNTR PCR
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DNA – coding versus non-coding regions Coding regions – DNA gene sequence that transcribes into protein (exons) Non-coding regions (95-98% of human DNA): – DNA that does not contain information for making proteins. – some non-coding DNA may be involved in regulation of transcription, translation, or gene expression during development. L. Mclane and A. Escobar
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A comparison of mutation frequency coding vs. non-coding regions Which line represents coding regions, and which line represents non-coding regions? Think about which has more mutations! L. Mclane and A. Escobar
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Review: S hort T andem R epeats (STR) regions in DNA of repeated sequences of 2-4 base pairs that repeat between 5-15 times inherited regions of non-coding DNA that vary from person to person STR polymorphisms are due to the different number of copies of the repeat element that occur in a population of individuals European Initiative for Biotechnology Profil
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ariable N umber of T andem R epeats (VNTR) regions of DNA of repeated sequences of 20- 50 base pairs that repeat from 50 to several hundred times the number of repeats is inherited according to the Mendelian principle of segregation in genetic terms a VNTR region is a locus with
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Bio 142 - Week 3 slides - Sp12-4 - PCR and Bioinformatics:...

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