![Page 1: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/1.jpg)
work summary
Wangqian
2007-01-12
![Page 2: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/2.jpg)
Mix samples(ARC1258 and ARC1218)
0/100 1/99 5/95 10/90 50/50 90/10 95/5 99/1 100/0
1258A.franciscana
0mg 0.6mg 2.6mg 5mg 25mg 45mg 47.4mg 49.5mg 50mg
1218A.sinica
50mg 49.4mg
47.6mg 45.2mg 25mg 5mg 2.5mg 6mg 0mg
total 50mg 50mg 50.2mg 50.2mg 50mg 50mg 49.9mg 50.1mg 50mg
![Page 3: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/3.jpg)
• 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.
![Page 4: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/4.jpg)
• 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 ℃
![Page 5: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/5.jpg)
Amplification (the kit of Taq DNA Polymerase)
item Final concentration
Tris-HCl(pH 8.3) 10mM
Mg2+ 2.5mM
dNTP 0.2mM
primers Each 0.25uM
BAS 1x
Taq 0.2u
DNA Dilution 1000 times DNA, 5ul
Total 50ul
![Page 6: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/6.jpg)
PCR (0/100 and 100/0)
• Dilute DNA(20ng/ul) 0 time, 10times, 100times, 1000times.
• Amplication 30cycle, 28cycle, 26cycle.
![Page 7: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/7.jpg)
![Page 8: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/8.jpg)
![Page 9: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/9.jpg)
Count the cyst with magnifier
0/100 1/99 5/95 10/90 50/50 90/10 95/5 99/1 100/0
1258A.franciscana
0 1 5 10 50 90 95 99 100
1218A.sinica
100 99 95 90 50 10 5 1 0
total 100 100 100 100 100 100 100 100 100
![Page 10: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/10.jpg)
• 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).
![Page 11: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/11.jpg)
![Page 12: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/12.jpg)
• 0/100 and 100/0
• Use 3ul of supernatant for 50ul PCR reaction.
• 26cycle,28cycle,30cycle,32cycle,34cycle,36cycle.
DNA extracted by chelex 6%
![Page 13: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/13.jpg)
![Page 14: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/14.jpg)
Promega and chelex 6%
![Page 15: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/15.jpg)
![Page 16: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/16.jpg)
![Page 17: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/17.jpg)
• 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
![Page 18: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/18.jpg)
![Page 19: Work summary Wangqian 2007-01-12. Mix samples(ARC1258 and ARC1218) 0/1001/995/9510/9050/5090/1095/599/1100/0 1258 A.franc iscana 0mg0.6mg2.6mg5mg25mg45mg47.4mg49.5mg50mg](https://reader034.vdocuments.site/reader034/viewer/2022051619/56649d595503460f94a39763/html5/thumbnails/19.jpg)