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演示文稿演讲PPT学习教学课件医学文件教学培训课件
Training on STR Typing Using Commercial Kits and ABI 310/3100 Margaret C. Kline, Janette W. Redman, John M. Butler National Institute of Standards and Technology October 22-26, 2001 Human Identity Testing Forensic cases -- matching suspect with evidence Paternity testing -- identifying father Historical investigations Missing persons investigations Mass disasters -- putting pieces back together Military DNA “dog tag” Convicted felon DNA databases DNA Quantitation using Slot Blot AMEL D3 TH01 TPOX Penta D Penta E FGA D21 D18 CSF D16 D7 D13 D5 VWA D8 PCR Amplification with Fluorescent STR Kits and Separation with Capillary Electrophoresis Blood Stain Overview of Steps Involved in DNA Typing Genotyping by Comparison to Allelic Ladder Sample Obtained from Crime Scene or Paternity Investigation DNA Extraction DNA Quantitation PCR Amplification of Multiple STR markers Biology Separation and Detection of PCR Products (STR Alleles) Technology Sample Genotype Determination Genetics Comparison of Sample Genotype to Other Sample Results If match occurs, comparison of DNA profile to population databases Generation of Case Report with Probability of Random Match Steps in Sample Processing Calculation of DNA Quantities in Genomic DNA Important values for calculations: 1 bp = 618 g/mol A: 313 g/mol; T: 304 g/mol; A-T base pairs = 617 g/mol G: 329 g/mol; C: 289 g/mol; G-C base pairs = 618 g/mol ? 1 genome copy = ~3 x 109 bp = 23 chromosomes (one member of each pair) ? 1 mole = 6.02 x 1023 molecules ? Standard DNA typing protocols with PCR amplification of STR markers typically ask for 1 ng of DNA template. How many actual copies of each STR locus exist in 1 ng? ? ? 1 genome copy = (~3 x 109 bp) x (618 g/mol/bp) = 1.85 x 1012 g/mol ? = (1.85 x 1012 g/mol) x (1 mole/6.02 x 1023 molecules) ? = 3.08 x 10-12 g = 3.08 picograms (pg) ? ? Since a diploid human cell contains two copies of each chromosome, then ? each diploid human cell contains ~6 pg genomic DNA ? ? 1 ng gen
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