Team:NTNU Trondheim/Notebook

From 2012.igem.org

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<h1>Notebook <small>Unfiltered notes from the lab</small></h1>
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__NOTOC__
__NOTOC__
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=Notebook=
 
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Click on a month name or date to view the respective notebook entries. For information on plasmids and DNA mentioned in the notebook, see [http://2012.igem.org/Team:NTNU_Trondheim/Stocks Stocks] and [http://2012.igem.org/Team:NTNU_Trondheim/Constructs Constructs]. For explanations of abbreviations used, click [http://2012.igem.org/Team:NTNU_Trondheim/Abbreviations here].
Click on a month name or date to view the respective notebook entries. For information on plasmids and DNA mentioned in the notebook, see [http://2012.igem.org/Team:NTNU_Trondheim/Stocks Stocks] and [http://2012.igem.org/Team:NTNU_Trondheim/Constructs Constructs]. For explanations of abbreviations used, click [http://2012.igem.org/Team:NTNU_Trondheim/Abbreviations here].
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Did a restriction digest as described in the protocol on the following parts.
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Did a restriction digest as described in the [http://2012.igem.org/Team:NTNU_Trondheim/Protocols protocol] on the following parts. On the pieces to be used as inserts, the number of base pairs are also listed.
{| class="wikitable" style="text-align:center;margin: 1em auto 1em auto;"
{| class="wikitable" style="text-align:center;margin: 1em auto 1em auto;"
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!Enzymes
!Enzymes
!Buffer
!Buffer
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!bp insert
|-
|-
|LuxI+term
|LuxI+term
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|XbaI+PstI
|XbaI+PstI
|3
|3
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|747
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|-
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|YFP+term
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|<partinfo>BBa_E0030</partinfo>, <partinfo>BBa_B0015</partinfo>
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|XbaI+PstI
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|3
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|852
|-
|-
|Lysis
|Lysis
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|XbaI+PstI
|XbaI+PstI
|3
|3
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|1785
|-
|-
|LuxI
|LuxI
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|XbaI+PstI
|XbaI+PstI
|3
|3
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|618
|-
|-
|RBS*
|RBS*
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|SpeI+PstI
|SpeI+PstI
|1
|1
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| -
|-
|-
|P<sub>luxR+HSL</sub>
|P<sub>luxR+HSL</sub>
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|SpeI+PstI
|SpeI+PstI
|1
|1
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| -
|-
|-
|RBS
|RBS
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|SpeI+PstI
|SpeI+PstI
|1
|1
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| -
|-
|-
|pSB1A3
|pSB1A3
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|EcoRI+PstI+DpnI
|EcoRI+PstI+DpnI
|3
|3
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| -
|-
|-
|plld
|plld
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|EcoRI+SpeI
|EcoRI+SpeI
|4
|4
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|433
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The parts lysis <partinfo>BBa_K112808</partinfo>, YFP+term (<partinfo>BBa_E0030</partinfo>, <partinfo>BBa_B0015</partinfo>), <partinfo>pSB1A3</partinfo> and plld will be assembled using the 3A assembly, so made double cutting mix with these parts.
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Gel electrophoresis was done with luxI+term (<partinfo>Bba_C0061</partinfo>, <partinfo>BBa_B0015</partinfo>), lysis <partinfo>BBa_K112808</partinfo>, YFP+term (<partinfo>BBa_E0030</partinfo>, <partinfo>BBa_B0015</partinfo>), LuxI <partinfo>Bba_C0061</partinfo> and plld.
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P<sub>lld</sub>+lysis and pSB1A3 cut yesterday with E+P was investigated on gel. Two fragments were obtained for P<sub>lld</sub>+lysis, but the sizes of the fragments were not as expected. Since we are not sure which fragment is which, both of the fragments were cut out of the gel and purified using the QIAquick gel extraction kit. In case one of them is uncut plasmid, the unligated DNA from both bonds will be transformed tomorrow, and we will assume that if the cells transformed with DNA from one of the samples won't grow, that sample will contain digested plasmid. This sample will be ligated with pSB1A3, plated out on a ampicillin plate, and then, several cell cultures from this plate will be plated out on a chloramphenicol plate. The colonies that won't grow on chloramphenicol, is assumed to contain the correct plasmid.
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Vgb+RBS religated, Vgb+RBS+LuxR+DTT #1, Vgb+RBS+LuxR+DTT #2, LuxI+DTT, K+RBS+LacI+DTT and his-tagged LldR was miniprepped using the Promega SV miniprep kit. The concentrations of isolated plasmid are given below:
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{| class="wikitable" style="text-align:center;margin: 1em auto 1em auto;"
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!Plasmid
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!Concentration [ng/µl]
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|-
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|Vgb+RBS religated
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|27.3
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|-
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|Vgb+RBS+LuxR+DTT #1
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|26.2
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|-
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|Vgb+RBS+LuxR+DTT #2
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|31.7
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|-
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|LuxI+DTT
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|35.3
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|-
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|K+RBS+LacI+DTT
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|7.9
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|-
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|his-LldR
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|51.0
|}
|}
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The parts lysis, YFP+term, pSB1A3 and plld will be assembled using the 3A assembly, made double cutting mix of these.
 
===Wednesday 08.08.12===
===Wednesday 08.08.12===
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We recieved emails from Eivind letting us know that we are the ones that have been selected to represent NTNU in iGEM 2012. Everybody is happy!
We recieved emails from Eivind letting us know that we are the ones that have been selected to represent NTNU in iGEM 2012. Everybody is happy!
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Latest revision as of 17:15, 25 September 2012

NTNU IS B.A.C.K.
Bacterial Anti-Cancer-Kamikaze

Notebook Unfiltered notes from the lab


February
Mon Tue Wed Thu Fri Sat Sun
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 25
27 28 29
March
Mon Tue Wed Thu Fri Sat Sun
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
April
Mon Tue Wed Thu Fri Sat Sun
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
May
Mon Tue Wed Thu Fri Sat Sun
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
June
Mon Tue Wed Thu Fri Sat Sun
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
July
Mon Tue Wed Thu Fri Sat Sun
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
August
Mon Tue Wed Thu Fri Sat Sun
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
Mon Tue Wed Thu Fri Sat Sun
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

Click on a month name or date to view the respective notebook entries. For information on plasmids and DNA mentioned in the notebook, see Stocks and Constructs. For explanations of abbreviations used, click here.



Retrieved from "http://2012.igem.org/Team:NTNU_Trondheim/Notebook"