Team:Kyoto/GSDD/Notebook/0831-0906

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Construction.
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Construction.
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Construction.PCR.
0824-0830
Construction.PCR.
0831-0906
Construction.PCR.MPR.
0907-0913
Construction.PCR.MPR.
0914-0920
Construction.PCR.MPR.
0921-0927
Construction.PCR.MPR.
0928-1004
Construction.PCR.MPR.
1005-1011
Construction.PCR.MPR.
1012-1018
Construction.MPR.Observation.

0831-0906 : Construction.PCR.MPR.


Monday, 31 August

To Do

  • Miniprep p2-1-1, p2-1-2, p2-4-2
  • Transformation p2-4-1(retry)
  • MPR

Results

Miniprep
  • p2-1-1, p2-1-2, p2-4-2 was minipreped.
Miniprep conc.
NO.Sample nameconc./ul
1p2-1-167.9
2p2-1-261.5
3p2-4-294.4
Transformation
  • p2-4-1 was transformed, and colony was observed.
MPR
  • xxx was MPRed.
18 MPR samples
NO.123456
temp.X deg C606060606060
MPR primer 1Fw1Fw2Fw3Fw1Fw2Fw3
MPR primer 2Rev1Rev1Rev1Rev2Rev2Rev2
NO.123456
temp.X deg C636363636363
MPR primer 1Fw1Fw2Fw3Fw1Fw2Fw3
MPR primer 2Rev1Rev1Rev1Rev2Rev2Rev2
NO.123456
temp.X deg C666666666666
MPR primer 1Fw1Fw2Fw3Fw1Fw2Fw3
MPR primer 2Rev1Rev1Rev1Rev2Rev2Rev2

Kyoto MPR 0831.png

  • Bands were observed on samples with Fw2.
  • Bands were longer on samples with Rev1 than with Rev2.



Tuesday, 1 September

To Do

  • Electrophoresis of MPR produnts on 31 Aug.
  • Miniprep

Results

Electrophoresis

(See lab-note on 31 Aug.)



Wednesday, 2 September

To Do

  • Miniprep of p2-4-1
  • PCR of (7), (8), (10)
  • PCR of (7), (10) (retry)

Results

Miniprep
  • p2-4-1 was minipreped.
Miniprep conc.
NO.Sample nameconc./ul
1p2-4-192.1
PCR
  • (7), (8), (10) was PCRed.
PCR conc.
NO.Sample nameconc./(ng/ul)
1(7)9.7
2(8)73.2
3(10)6.9
  • (7), (10) was PCRed twice (again).
PCR conc.
NO.Sample nameconc./(ng/ul)
1(7)A2.9
2(7)B9.3
3(10)A5.5
4(10)B9.1

These concentration suggest that PCR was not achieved. Low concentration of primer causes PCR fault. We should have mix well primer when we add water in primer tube. We are going to retry PCR of (7),(10) the next day.



Thursday, 3 September

To Do

  • measuring concentration of (7), (10) (see lab-note on 2 Sept.)
  • PCR and measuring concentration of (7), (10) (retry)
  • Restriction Enzyme, and PCR purification (8).

Results

Electrophoresis of PCR (7), (10)

Kyoto PCR 0903.png

Concentration of PCR (7), (10)
PCRed conc.
NO.Sample nameconc./(ng/ul)
1(7)A71.7
2(7)B104.3
3(10)A33.5
4(10)B42.6



Friday, 4 September

To Do

(5)

  • PCR
  • PCR purification kit

(5), (7), (10)

  • Restriction Enzyme
  • purification kit
  • ligation

Results

Electrophoresis of PCR (5)

Kyoto PCR 0904.png

Concentration of PCR (5)
PCRed conc.
NO.Sample nameconc./(ng/ul)
1(5)A143.4
2(5)B174.8
Concentration of PCR prification kit (5), (7), (8), (10)
PCR prified conc.
NO.Sample nameconc./(ng/ul)
1(5)A53.9
2(5)B95.4
3(7)A28.9
4(7)B33.3
5(8)68.3
6(10)A29.0
7(10)B32.2