Team:TUDelft/5 August 2009

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='''5 August 2009'''=
='''5 August 2009'''=
 +
===Sriram===
 +
Today I did the restiction along with Daniel. After lot of calculation and discussion we did it successfully. Then I prepared a gel and loaded all the restricted samples in the gel and it could be seen in below image:
 +
 +
[[Image:igemdelay060809.jpg|thumb|center|450px]]
 +
 +
Then we decided to continue the assembly tomorrow. Daniel gave me the instruction and he decided to check with how to do the GFP expression experiments.
===Tim Weenink===
===Tim Weenink===
-
[[Image:Tim050809.png]]
+
[[Image:Tim050809.png|center|400px]]
 +
 
 +
 
 +
[[Image:Tim050809!Dandcalin.png|center|400px]]
 +
 
 +
 
 +
{| border="1" align="center"
 +
| Well || Part || Expected Plasmid Size || Status
 +
|- align="center"
 +
| 1-8 || !D PCR products (wrong primer) || 1100 ||  <font color=red>&#10006;</font>
 +
|- align="center"
 +
| 9 || [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder] ||  ||  <font color=limegreen>&#10004;</font>
 +
|- align="center"
 +
| 10 || pSB1AK3 linear || 3864 ||  <font color=limegreen>&#10004;</font>
 +
|- align="center"
 +
| 11 || pSB4C5 linear || 3896 || <font color=limegreen>&#10004;</font>
 +
|- align="center"
 +
| 12 || strong promoter || 2983 || <font color=limegreen>&#10004;</font>
 +
|- align="center"
 +
| 13 || oriT-R linear || ? || <font color=limegreen>&#10004;</font>
 +
|}
 +
 
 +
===Orr===
 +
Prepared 1 liter of LB medium, and helped Calin with the Conjugation procedure.
 +
 
 +
===Calin===
 +
 
 +
Made test plate for Trimpethoprim.
 +
 
 +
Ran conjugation test with R751 (TRI) as donor and DH5 &alpha; cells with GFP plasmid and AMP as recipients.
 +
 
 +
Ran three conjugations in parallel, with a conjugation time of 1 hour.
 +
 
 +
Starting OD for recipients 0.502, for donors 0.542.
 +
 
 +
Made a total of 40 plates:
 +
 
 +
 
 +
{| border="1" align="center"
 +
|-
 +
| <b>Set 1</b>
 +
|- align="center"
 +
| Plate ID || Antibiotics || Dilution ||
 +
|- align="center"
 +
| D1 || TRI || 10<sup>0</sup> ||
 +
|- align="center"
 +
| D2 || TRI || 10<sup>-2</sup>||
 +
|- align="center"
 +
| D3 || TRI || 10<sup>-4</sup>||
 +
|- align="center"
 +
| D4 || TRI || 10<sup>-5</sup>||
 +
|- align="center"
 +
| D5 || TRI || 10<sup>-6</sup>||
 +
|- align="center"
 +
| T1 || TRI + AMP || 10<sup>0</sup> ||
 +
|- align="center"
 +
| T2 || TRI + AMP || 10<sup>-1</sup>||
 +
|- align="center"
 +
| T3 || TRI + AMP || 10<sup>-2</sup>||
 +
|- align="center"
 +
| T4 || TRI + AMP || 10<sup>-3</sup>||
 +
|- align="center"
 +
| T5 || TRI + AMP || 10<sup>-4</sup>||
 +
|- align="center"
 +
| T6 || TRI + AMP || 10<sup>-5</sup> ||
 +
|- align="center"
 +
| T7 || TRI + AMP || 10<sup>-6</sup>||
 +
|- align="center"
 +
| R1 || AMP || 10<sup>0</sup> ||
 +
|- align="center"
 +
| R2 || AMP || 10<sup>-4</sup>||
 +
|- align="center"
 +
| R3 || AMP || 10<sup>-5</sup>||
 +
|- align="center"
 +
| R4 || AMP || 10<sup>-6</sup>||
 +
|}
 +
 
 +
<br><br>
 +
{| border="1" align="center"
 +
|-
 +
| <b>Set 2</b>
 +
|- align="center"
 +
| Plate ID || Antibiotics || Dilution ||
 +
|- align="center"
 +
| D6 || TRI || 10<sup>0</sup> ||
 +
|- align="center"
 +
| D7 || TRI || 10<sup>-2</sup>||
 +
|- align="center"
 +
| D8 || TRI || 10<sup>-4</sup>||
 +
|- align="center"
 +
| D9 || TRI || 10<sup>-5</sup>||
 +
|- align="center"
 +
| D10 || TRI || 10<sup>-6</sup>||
 +
|- align="center"
 +
| T8 || TRI + AMP || 10<sup>0</sup> ||
 +
|- align="center"
 +
| T9 || TRI + AMP || 10<sup>-1</sup>||
 +
|- align="center"
 +
| T10 || TRI + AMP || 10<sup>-2</sup>||
 +
|- align="center"
 +
| T11 || TRI + AMP || 10<sup>-3</sup>||
 +
|- align="center"
 +
| T12 || TRI + AMP || 10<sup>-4</sup>||
 +
|- align="center"
 +
| T13 || TRI + AMP || 10<sup>-5</sup> ||
 +
|- align="center"
 +
| T14 || TRI + AMP || 10<sup>-6</sup>||
 +
|}
 +
<br><br>
 +
{| border="1" align="center"
 +
|-
 +
| <b>Set 3</b>
 +
|- align="center"
 +
| Plate ID || Antibiotics || Dilution ||
 +
|- align="center"
 +
| D11 || TRI || 10<sup>0</sup> ||
 +
|- align="center"
 +
| D12 || TRI || 10<sup>-2</sup>||
 +
|- align="center"
 +
| D13 || TRI || 10<sup>-4</sup>||
 +
|- align="center"
 +
| D14 || TRI || 10<sup>-5</sup>||
 +
|- align="center"
 +
| D15 || TRI || 10<sup>-6</sup>||
 +
|- align="center"
 +
| T15 || TRI + AMP || 10<sup>0</sup> ||
 +
|- align="center"
 +
| T16 || TRI + AMP || 10<sup>-1</sup>||
 +
|- align="center"
 +
| T17 || TRI + AMP || 10<sup>-2</sup>||
 +
|- align="center"
 +
| T18 || TRI + AMP || 10<sup>-3</sup>||
 +
|- align="center"
 +
| T19 || TRI + AMP || 10<sup>-4</sup>||
 +
|- align="center"
 +
| T20 || TRI + AMP || 10<sup>-5</sup> ||
 +
|- align="center"
 +
| T21 || TRI + AMP || 10<sup>-6</sup>||
 +
|}
{{Template:TUDelftiGEM2009_end}}
{{Template:TUDelftiGEM2009_end}}

Latest revision as of 08:40, 20 October 2009

Lab Notebook

July
MTWTFSS
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6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31
August
MTWTFSS
          1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31
September
MTWTFSS
  1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30
October
MTWTFSS
      1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30 31
 

5 August 2009

Sriram

Today I did the restiction along with Daniel. After lot of calculation and discussion we did it successfully. Then I prepared a gel and loaded all the restricted samples in the gel and it could be seen in below image:

Igemdelay060809.jpg

Then we decided to continue the assembly tomorrow. Daniel gave me the instruction and he decided to check with how to do the GFP expression experiments.

Tim Weenink

Tim050809.png


Tim050809!Dandcalin.png


Well Part Expected Plasmid Size Status
1-8  !D PCR products (wrong primer) 1100 ✖
9 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder] ✔
10 pSB1AK3 linear 3864 ✔
11 pSB4C5 linear 3896 ✔
12 strong promoter 2983 ✔
13 oriT-R linear  ? ✔

Orr

Prepared 1 liter of LB medium, and helped Calin with the Conjugation procedure.

Calin

Made test plate for Trimpethoprim.

Ran conjugation test with R751 (TRI) as donor and DH5 α cells with GFP plasmid and AMP as recipients.

Ran three conjugations in parallel, with a conjugation time of 1 hour.

Starting OD for recipients 0.502, for donors 0.542.

Made a total of 40 plates:


Set 1
Plate ID Antibiotics Dilution
D1 TRI 100
D2 TRI 10-2
D3 TRI 10-4
D4 TRI 10-5
D5 TRI 10-6
T1 TRI + AMP 100
T2 TRI + AMP 10-1
T3 TRI + AMP 10-2
T4 TRI + AMP 10-3
T5 TRI + AMP 10-4
T6 TRI + AMP 10-5
T7 TRI + AMP 10-6
R1 AMP 100
R2 AMP 10-4
R3 AMP 10-5
R4 AMP 10-6



Set 2
Plate ID Antibiotics Dilution
D6 TRI 100
D7 TRI 10-2
D8 TRI 10-4
D9 TRI 10-5
D10 TRI 10-6
T8 TRI + AMP 100
T9 TRI + AMP 10-1
T10 TRI + AMP 10-2
T11 TRI + AMP 10-3
T12 TRI + AMP 10-4
T13 TRI + AMP 10-5
T14 TRI + AMP 10-6



Set 3
Plate ID Antibiotics Dilution
D11 TRI 100
D12 TRI 10-2
D13 TRI 10-4
D14 TRI 10-5
D15 TRI 10-6
T15 TRI + AMP 100
T16 TRI + AMP 10-1
T17 TRI + AMP 10-2
T18 TRI + AMP 10-3
T19 TRI + AMP 10-4
T20 TRI + AMP 10-5
T21 TRI + AMP 10-6

Retrieved from "http://2009.igem.org/Team:TUDelft/5_August_2009"