Team:Osaka/week6

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Contents

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
NameIPTG(0.1M)incubationOD600A474
1 --0.85830.1567
2400μl2h1.15230.1447
3400μl1h0.55830.1126
440μl2h1.02970.1878
540μl1h0.65760.1736
  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 of ligation products
  11. Absorbance measurement
Table 8
NameUV irradiationincubationOD600A474
03① --
03②1mJ/cm21h
04① --
04②1mJ/cm21h

September 10(Sat)

  1. PCR
  2. Electrophoresis of PCR products
  3. To liquid culture:101,102
  4. PCR
  5. PCR purification
  6. Absorbance measurement
Table 9
NameUV irradiationincubationOD600A474
03① --
03②1mJ/cm21h
04① --
04②1mJ/cm21h


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