Team:Osaka/week6

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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)

1. PCR
2. Electrophoresis of PCR products
3. Gel extraction
4. Restriction digests
5. PCR
6. Electrophoresis of PCR products
7. Gel extraction
8. Restriction digests
9. Ligation
   • 101(C):PprI+1-3A(vector)
   • 102(C):PprA+1-3A(vector)
10. Transformation
11. Absorbance measurement

Table 8

Name	UV irradiation	incubation	OD600	A474
03①	-	-
03②	1mJ/cm2	1h
04①	-	-
04②	1mJ/cm2	1h

September 10(Sat)

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