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work summary

work summary. Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218). DNA extraction : use the Wizard Genomic DNA Purification kit (Promega TM , mouse tail protocol) DNA dilute 20 times to PCR, then DNA concentration is unified to 20ng/ul. The primers are 12S-SP and 12S-SF-GC

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work summary

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  1. work summary Wangqian 2007-01-12

  2. Mix samples(ARC1258 and ARC1218)

  3. DNA extraction : use the Wizard Genomic DNA Purification kit (Promega TM, mouse tail protocol) • DNA dilute 20 times to PCR, then DNA concentration is unified to 20ng/ul.

  4. The primers are 12S-SP and 12S-SF-GC 12S-SP: 5’-CTA-GGA-TTA-GAT-ACC-CTA-3’ 12S-SF-GC: 5’-CCG-GGG-CCC-GCG-GGC-CCC-CGG-GCC-GGG-CCC-GGG-GAG-AGC-GAC-GGG-CGT-ATG-TAT-3’ • PCR condition: 94℃, 2m, 1 cycle 26x, 28x, 30x94℃, 30s; 51℃,30s; 72℃,1m, 72℃, 4m , 1 cycle

  5. Amplification (the kit of Taq DNA Polymerase)

  6. PCR (0/100 and 100/0) • Dilute DNA(20ng/ul) 0 time, 10times, 100times, 1000times. • Amplication 30cycle, 28cycle, 26cycle.

  7. Count the cyst with magnifier

  8. DNA extraction : use the Wizard Genomic DNA Purification kit (Promega TM, mouse tail protocol) • DNA concentration is 10ng/ul. • Count cyst again, extract the DNA with Chelex 6%(Bio-Rod).

  9. DNA extracted by chelex 6% • 0/100 and 100/0 • Use 3ul of supernatant for 50ul PCR reaction. • 26cycle,28cycle,30cycle,32cycle,34cycle,36cycle.

  10. Promega and chelex 6%

  11. use samples from ghent and dvz • Mix by 50/50 • Extract DNA by promage, concentration of DNA are 9.5ng/ul and 21ng/ul. then dilute DNA to 9.5ng/ul. • Dilute DNA 1000times to PCR • 32cycle

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