1 / 19

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

jessica-jarvis
Download Presentation

work summary

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  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

More Related