Team:Osaka/week6

Contents 1 September 4(Sun) 2 September 5(Mon) 3 September 6(Tue) 4 September 7(Wed) 5 September 8(Thu) 6 September 9(Fri) 7 September 10(Sat)

September 4(Sun)

1. Restriction digests:3-10C,1-2M
2. Gel extraction of digests
3. Ligation
   • 03:1-9N+3-6H again
4. Transformation of ligation products
5. Photospectrometry measuament

September 5(Mon)

1. Absorbance measurement

Table 7

Name	IPTG(0.1M)	incubation	OD600	A474
1	-	-	0.8583	0.1567
2	400μl	2h	1.1523	0.1447
3	400μl	1h	0.5583	0.1126
4	40μl	2h	1.0297	0.1878
5	40μl	1h	0.6576	0.1736

1. To liquid culture:03,17,3-10C

September 6(Tue)

1. Miniprep of yesterday's culture
2. Restriction digests of minipreppped parts
3. Gel electrophoresis of digests
   • all parts ×
4. PCR to amplify PprI,PprA,PprM,RecA (DNA repair protein) from D.radiodurance
5. Gel Electrophoresis of PCR products
   • PprM ok
   • PprI,PprA,RecA ×
6. PCR
7. Gel Electrophoresis of PCR products
   • ×

September 7(Wed)

1. PCR
2. Gel electrophoresis of PCR products
   • PprM ok
   • PprI,PprA,RecA ×
3. Gel electrophoresis of PprM
4. Gel extraction
5. PCR
6. Gel electrophoresis of PCR products
   • PprI,PprA,RecA ×
7. Miniprep of yesterday's culture
8. Restriction digests of miniprepped parts
9. Gel electrophoresis of digests
   • 03 ok
10. To liquid culture:03,04 (preincubation)
11. PCR
12. Electrophoresis of PCR products
13. PCR

September 8(Thu)

1. Electrophoresis of PCR products
2. Restriction digests of 1-3A(backbone)
3. PCR
4. Electrophoresis of PCR products and miniprepped parts
5. Gel extraction of 1-3A
6. PCR
7. Electrophoresis of PCR products
8. Inoculation of 03,04
9. To liquid culture 03,04
10. PCR
11. Electrophoresis of PCR products

September 9(Fri)

September 10(Sat)

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