Team:TUDelft/18 August 2009

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Lab Notebook

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18 August 2009

Sriram

Checking the yesterday's gel (shown below) I found that since the ladder didn't run well we couldn't interpret the size properly. hence I decided to do the whole assembly again. Or else we need to follow the back up plan for negative cascade alone created by Daniel.

Calin

Yesterdays 20 well 1% gel:

Gel17082009-20well-1pc-sd-c.png


Well Part Expected Plasmid Size Status
1
2
3
4
5
6 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder] ✔
7 Colony PCR 1A - R751ΔtrbK 1616 ✖
8 Colony PCR 1A - R751ΔtrbK 1616 ✖
9 Colony PCR R751 + pKD46 + trbK_KO_mod_S 348 ✔
10 Colony PCR R751ΔoriTR -L-ara +PCR control  ?
11 Colony PCR R751 + pKD46 + oriTR_KO_mod_S 398 ✔
12 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder] ✔

Yesterdays 26 well 2% gel:

Gel17082009-26well-2pc-dela.png

Well Part Expected Plasmid Size Status
1 pTet
2 RBS-cI-RBS
3 pSB1C3
4 pLacI
5 RBS
6 pSB1C3
7 pTet
8 lock3c
9 pSB1C3 (not loaded well)
10 pSB1C3
11 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder] ✔
12 cI
13 Double Terminator
14 pSB1C3
15 λp-in
16 RBS-GFP-Double Term
17 pSB1C3
18 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder] ✖
19 pLacI
20 key3c
21 pSB1AK3
22 Colony PCR 1A - R751ΔtrbK 1616 ✖
23 Colony PCR 1A - R751ΔtrbK 1616 ✖
24 Colony PCR R751 + pKD46 + trbK_KO_mod_S 348 ✔
25 Colony PCR R751ΔoriTR -L-ara +PCR control  ?
26 Colony PCR R751 + pKD46 + oriTR_KO_mod_S 398 ✔


For the trbK conjugation test 24 plates were made:


Set 1
Plate ID Antibiotics Dilution
D1tr TRI 100
D2tr TRI 10-1
D3tr TRI 10-2
D4tr TRI 10-3
D5tr TRI 10-4
T1tr TRI + CAM 100
T2tr TRI + CAM 10-1
T3tr TRI + CAM 10-2
T4tr TRI + CAM 10-3
T5tr TRI + CAM 10-4
R1tr CAM 100
R2tr CAM 10-1



Set 2
Plate ID Antibiotics Dilution
D6tr TRI 100
D7tr TRI 10-1
D8tr TRI 10-2
D9tr TRI 10-3
D10tr TRI 10-4
T6tr TRI + CAM 100
T7tr TRI + CAM 10-1
T8tr TRI + CAM 10-2
T9tr TRI + CAM 10-3
T10tr TRI + CAM 10-4
R3tr CAM 100
R4tr CAM 10-1

For the oriTR conjugation test 51 plates were made:

Set 1
Plate ID Antibiotics Dilution
D1ori TRI + CAM 100
D2ori TRI + CAM 10-1
D3ori TRI + CAM 10-2
D4ori TRI + CAM 10-3
D5ori TRI + CAM 10-4
Ts1ori AMP + CAM 100
Ts2ori AMP + CAM 10-1
Ts3ori AMP + CAM 10-2
Ts4ori AMP + CAM 10-3
Ts5ori AMP + CAM 10-4
TL1ori TRI + AMP 100
TL2ori TRI + AMP 10-1
TL3ori TRI + AMP 10-2
TL4ori TRI + AMP 10-3
TL5ori TRI + AMP 10-4
R1ori AMP 100
R2ori AMP 10-1



Set 2
Plate ID Antibiotics Dilution
D6ori TRI + CAM 100
D7ori TRI + CAM 10-1
D8ori TRI + CAM 10-2
D9ori TRI + CAM 10-3
D10ori TRI + CAM 10-4
Ts6ori AMP + CAM 100
Ts7ori AMP + CAM 10-1
Ts8ori AMP + CAM 10-2
Ts9ori AMP + CAM 10-3
Ts10ori AMP + CAM 10-4
TL6ori TRI + AMP 100
TL7ori TRI + AMP 10-1
TL8ori TRI + AMP 10-2
TL9ori TRI + AMP 10-3
TL10ori TRI + AMP 10-4
R3ori AMP 100
R4ori AMP 10-1




Set 3
Plate ID Antibiotics Dilution
D11ori TRI + CAM 100
D12ori TRI + CAM 10-1
D13ori TRI + CAM 10-2
D14ori TRI + CAM 10-3
D15ori TRI + CAM 10-4
Ts11ori AMP + CAM 100
Ts12ori AMP + CAM 10-1
Ts13ori AMP + CAM 10-2
Ts14ori AMP + CAM 10-3
Ts15ori AMP + CAM 10-4
TL11ori TRI + AMP 100
TL12ori TRI + AMP 10-1
TL13ori TRI + AMP 10-2
TL14ori TRI + AMP 10-3
TL15ori TRI + AMP 10-4
R5ori AMP 100
R6ori AMP 10-1

Tim Weenink

Results of the 24h sequencing are in:

  • !BPEA1 (R) correct in reliable part of sequencing file
  • !BCEC1 (R) correct in reliable part of sequencing file
  • !CCEA1 (F) perfect
  • !CCEC2 (F) perfect
  • !SPEA1 (F) Assebly with !A and BBa_K142202. !A is not supposed to be there
  • !SPEA1 (R) Assebly with !A and BBa_K142202. !A is not supposed to be there
  • !SPEC1 (F) Failed (as expected)
  • !SPEC1 (R) Failed (as expected)
  • *I6F (F) Perfect
  • *S3 (F) Is not BBa_K142205, but in fact BBa_K142203
  • *S3 (R) Is not BBa_K142205, but in fact BBa_K142203

Daniel

Growth, miniprep:

Biobrick [DNA] ng/uL
C0051 45
P0440 48.2
R0051 22.8
I13504 70.1
Ca 28.1


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