Team:Toulouse/Notebook/project-monitoring

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Notebook > Project monitoring

Binding module

1. Amplification of Binding Module (pEX-K4) into E. coli

Transformation of Binding module (pEX-K4) into E. coli

Gene "Binding module" synthesized by Eurofins, resuspended in 20μL Tris 10mM
Result: We obtained distinct colonies on plates LB + Kanamycin (50 µg/mL)

Liquid culture of two clones: 1 et 2 (pEX-K4) transformed into E. coli

Date: 01/08/2014

Miniprep with QIAprep Spin Miniprep Kit: two clones of Binding Module (pEX-K4) into E. coli

Date: 04/08/2014
Result: Binding Module (pEX-K4) obtained

Digestion of Binding Module (pEX-K4) with EcoRI and PstI

Date: 04/08/2014
Result:
- 3 bands : 1500bp (vector with Kanamycin resistance), 1300bp (Module Binding) and 1000bp (vector with pUC Ori)
- The two Binding Module clones are ok

2. Cloning Binding Module on pEX-K4 with pSB1C3 (BBa_K606013 without RFP) into E. coli

Digestion Binding Module on pEX-K4 and BBa_K606013

Date: 23/08/2014
Expected bands after digestion:
- BBa_K606013: 860 bp for RFP and 2100 bp for vector pSB1C3
- Binding Module: 1400 bp for Binding Module and 1500bp + 1000bp for vector pEX_K4
We decide to do a ligation between pSB1C3 and Binding Module without gel cutting and purification but after a PCR clean-up (Thermo scientific, GeneJET PCR purifiction kit #K0701) to remove the enzymes.

Ligation Binding Module on pSB1C3

Date: 04/08/2014

Transformation of Binding Module on pSB1C3 into E. coli

Date: 04/08/2014
Transformation with 10 µL of the ligation mix and plate on Chloremphenicol LA plate
Result: many wrong clones

3. Cloning Binding Module on pEX-K4 with pVeg on pSB1C3 (BBa_K823003) into E. coli

Digestion Binding Module on pEX-K4 and BBa_K823003

Date: 04/08/2014
BBa_K823003 digested by XbaI and PstI and Binding Module digested by SpeIand PstI
Gel Electrophoresis
Result:
- Binding Module : 1400 bp for Binding Module and 1500bp + 1000bp for vector pEX_K47
We decide to do a ligation between pVeg and Binding Module without gel cutting and purification but after a PCR clean-up (Thermo scientific, GeneJET PCR purifiction kit #K0701) to remove the enzymes.

Ligation Binding Module on pVeg with pSB1C3

Date: 04/08/2014
BBa_K823003 digested by XbaI and PstI and Binding Module digested by SpeI and PstI and ligation

Transformation of Binding Module on pVeg into E. coli

Date: 04/08/2014
Result: Many good clones (check on 06/08/2014)

4. Cloning Binding Module with P_veg (BBa_K1364005) on pSB_BS4S (BBa_K823022) into E. coli

Digestion Binding Module with P_veg (BBa_K1364005) and pSB_BS4S (BBa_K823022)

Date: 07/08/2014
BBa_K823022 and Binding Module with P_veg (BBa_K1364005) digested by EcoRI and PstI Gel Electrophoresis
Result:
- Binding Module with Pveg 1600 bp for Binding Module and 2100bp for vector pSB1C3

Ligation Binding Module with Pveg on pSBbs4S

Date: 07/08/2014
BBa_K823022 and Binding Module with P_veg (BBa_K1364005) digested by EcoRI and PstI
Ligation
Result: Ligation between Binding Module with P_veg on pSB_BS4S

Transformation of Binding Module with P_veg on pSB_BS4S into E. coli

Date: 04/08/2014
Binding Module with P_veg on pSB_BS4S
Plate on Ampicillin LA plate
Result: Many good clones (check on 13/08/2014)

Transformation of Binding Module with P_veg on pSB_BS4S into B. subtilis

Date: 04/08/2014
After 5 hours of incubation and the linearization of 6µl of DNA with 1µl of ScaI, we make the transformation. Plate on Spectinomycin LA plate.
Result: One good clone : PCR (check on 09/09/2014) and threonine test (check on 09/09/2014)

5. Binding Test

See here

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Chemotaxis

1. Transformation of chemotaxis (Puc-57) into E. coli

Gene chemotaxis synthesized by Eurofins, resuspended in 20μL Tris 10mM
Date: 01/08/2014
Result: We obtained distinct colonies on LA + Ampicillin (100 µg/mL) plates

Culture of 4 clones: A, B, C, D of chemotaxis (Puc57) transformed into E. coli

Date: 04/08/2014

Miniprep with QIAprep Spin Miniprep Kit Using a Microcentrifuge: 4 clones of chemotaxis (Puc57) into E. coli

Date: 05/08/2014
Result: 4*50µL of chemotaxis (Puc57) obtained

2. Cloning chemotaxis BBa_K1364000 (chemotaxis_Puc57) with digested pSB1C3 with EcoRI and PstI into E. coli

Digestion BBa_K1364000 (chemotaxis_Puc57) with EcoRI and PstI - PCR kit Clean up

Date: 07/08/2014
Result: Expected band after digestion for BBa_K1364000: 2300 bp
Problem: We can't distinguish the vector band (2500 bp)

Gel extraction of BBa_K1364000

Date: 07/08/2014

Ligation BBa_K1364000 and digested PSB1C3 with EcoRI and PstI

Date: 08/08/2014

Transformation BBa_K1364000 in E.coli

Date: 08/08/2014
Result: We obtained distinct colonies on LA + Cm (15 µg/mL) plates and resuspended 15 colonies in LB+Cm (15 µg/mL)

Test of sensibility on Ampicillin

Date: 10/08/2014
Result: we can determine which colonies are sensible for ampicillin and know which bacterium is carrying the chemotaxis gene.

PCR

Date: 11/08/2014

Digestion BBa_K1364000 on pSB1C3 with EcoRI and PstI

Date: 11/08/2014
Result: There is one colony which presents the right construction.

3. Cloning chemotaxis BBa_K1364000 (chemotaxis_Puc57) with digested BBa_823003 (P_veg) on pSB1C3 with SpeI and PstI into E. coli

Ligation BBa_K1364000 and digested BBa_823003 on PsB1C3 with SpeI and PstI

Date: 08/08/2014

Transformation BBa_1364004 in E.coli

Date: 8/08/2014

Test of sensibility on Ampicillin

Date: 10/08/2014
Result: We can determine which colony is sensible for ampicillin and know which bacterium is carrying the chemotaxis gene. There is one colony which resists on ampicillin.

Digestion BBa_1364004 on pSBC3 with EcoRI and PstI

Date: 12/08/2014
Result: We did not see any colony with chemotaxis insert.

4. Cloning chemotaxis BBa_K1364000 (chemotaxis_Puc57) with digested BBa_823003 (P_veg) on pSB1C3 with SpeI and PstI into E. coli

Ligation BBa_K1364000 and digested BBa_823003 on PsB1C3 with SpeI and PstI

Date: 11/08/2014

Transformation BBa_1364004 in E.coli

Date: 11/08/2014

Test of sensibility on Ampicillin

Date: 14/08/2014
Result: we obtained 4 colonies sensible at Ampicilline

Digestion BBa_1364004 on PsB1C3 with EcoRI and PstI

Date: 18/08/2014

Gel extraction of BBa_1364000

Date: 19/08/2014

5. Cloning chemotaxis BBa_K1364004 with digested pSB_BS4S with EcorI and PstI into E. coli

Ligation BBa_K1364004 digested EcorI and PstI and digested PsB1C3 with EcoRI and PstI

Date: 19/08/2014

Transformation BBa_K1364004 in pSBBS4S in E.coli

Date: 19/08/2014
Result: We obtained one colony and resuspended it in LB+ Amp

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Fungicides

D4E1

1. Amplification of synthetic gene (D4E1 on pEX-A2)

Transformation of D4E1 (pEX-A2) into E. coli

Gene D4E1 synthesized by Eurofins, resuspended in 20μL Tris 10mM
Concentration of D4E1: 115ng/µL
Date: 07/21//2014
Result: We obtained distinct colonies on LA + Ampicillin (100 µg/mL)

Culture of 4 clones: A, B, C, D of D4E1 (pEX-A2) transformed into E. coli

Date: 07/22/2014
Result: Culture of 4 clones ok

QIAprep Spin Miniprep Kit Using a Microcentrifuge : 4 clones of D4E1 (pEX-A2) into E. coli

Buffer EB at 50-55°C
Date: 07/23/2014

Digestion of D4E1 (pEX-A2) with EcoRI and PstI - Stop EcoRI - PCR kit Clean up

Date: 07/23/2014
Result: 4*20µL D4E1 digested with EcoRI and PstI  all the clones seem to have the right D4E1 gene.

2. Cloning D4E1 in pSB1C3

Digestion of D4E1 on pEX-A2 and pSB1C3

Date: 07/23/2014

Ligation of D4E1 in pSB1C3

Date: 07/23/2014

Transformation in E.coli

Date: 07/23/2014

Culture of 6 clones: A, B, C, D, E, F of D4E1 (pSB1C3) transformed into E. coli

Processing details: 5mL of LB + chloramphenicol (15µg/mL) , using sterile plastic loop to culture colonies
Date: 07/24/2014
Result: Culture of 4 clones ok

QIAprep Spin Miniprep Kit Using a Microcentrifuge

Date: 07/25/2014

Digestion of D4E1 (pSB1C3) A, B, C, D, E, F with EcoRI and PstI - PCR kit Clean up + electrophoresis

Date: 07/25/2014
Result: clones C, D have the expected construction

PCR of D4E1 (pSB1C3) A, B, C, D, E, F + electrophoresis

Date: 07/25/2014
Result: clones C, D have the expected construction, and placed in cryopreservation.

3. Cloning Pveg+D4E1 on Pveg plasmid (pSB1C3)

Digestion of D4E1 on pEX-A2 and K823003 (Pveg on pSB1C3)

Dated: 07/24/2014
Result: 20µL digestion of D4E1 on pEX-A2 and of K823003

Ligation of D4E1 in K823003 (Pveg on pSB1C3)

Date: 07/24/2014
Result: 20µL ligation of D4E1 in K823003

Transformation of ligation products in E.coli

Date: 07/24/2014
Result: E.coli transformed by D4E1+K823003

Culture of 6 clones: A, B, C, D, E, F of transformed E. coli

Date: 07/26/2014
Result: Culture of 4 clones ok

QIAprep Spin Miniprep Kit Using a Microcentrifuge

Date: 07/28/2014

PCR of D4E1 (pSB1C3) A, B, C, D, E, F + electrophoresis

Dated: 07/28/2014
Result: clones E, F seem to have the expected construction

Digestion of clones E, F of D4E1+K823003 (Pveg on pSB1C3) with EcoRI and PstI + electrophoresis

Date: 28/07/2014
Result: clones C, D have the expected construction and are placed in cryopreservation.

4. Cloning P_veg + D4E1 on pSB_BS4S (K823022)

Date: 08/13/2014

5. Cloning P_veg + D4E1 on pSB_BS1C lacZ (23)

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GAFP1

1. Amplification of synthetic gene (GAFP1 on pEX-A2)

Transformation of GAFP1 (pEX-A2) into E.coli

Gene GAFP1 synthesized by Eurofins, resuspended in 20μL Tris 10mM
Concentration of GAFP1 : 145ng/µL
Date: 07/21/2014
Result: We obtained distinct colonies on plates LB + Ampicillin (100 µg/mL)

Culture of 4 clones: A, B, C, D of GAFP1 (pEX-A2) transformed into E. coli

Date: 07/22/2014
Result: Culture of 4 clones ok

QIAprep Spin Miniprep Kit Using a Microcentrifuge : 4 clones of GAFP1 (pEX-A2) into E. coli

Date: 07/23/2014

Digestion of GAFP1 (pEX-A2) with EcoRI and PstI - Inactivation EcoRI - PCR kit Clean up to remove PstI - Gel Electrophoresis

Date: 07/23/2014

2. Cloning GAFP1 gene on PSB1C3 = K1364002 in E. coli

Digestion GAFP1_pEX-A2 and BBa_K606013 (RFP_pSB1C3)

Date: 07/23/2014
Result: BBa_K606013 : 860 bp
We decide to conserve the miniprep B for BBa_K606013

Ligation GAFP1 and BBa_K606013

Date: 07/23/2014

Tranformation of ligation products into E. coli

Date: 07/23/2014

Culture of x clones of GAFP1+K606013 in E. coli

Date: 07/24/2014

QIAprep Spin Miniprep Kit Using a Microcentrifuge: 6 clones of GAFP1+K606013 = K1364002 (pSB1C3) into E. coli

Date: 07/25/2014

PCR on 6 clones of GAFP1+K606013 = K1364002 (pSB1C3) + electrophoresis

Date: 07/25/2014
Result: clones A, C, D, F seem to have the right construction

Digestion of 6 clones of GAFP1+K606013 = K1364002 (pSB1C3) by EcoR1 and Pst1 + electrophoresis

Date: 07/25/2014

3. Cloning GAFP1+terminator(B0015) = K1364007 (pSB1C3)

Digestion of GAFP1 on pEX-A2 and B0015 (terminator on pSB1C3)

Date: 07/24/2014

Ligation of GAFP1 in B0015 (terminator on pSB1C3)

Date: 07/24/2014

Transformation of ligation products in E.coli

Date: 07/24/2014

Culture of 6 clones: A, B, C, D, E, F transformed in E. coli

Date: 07/26/2014

QIAprep Spin Miniprep Kit Using a Microcentrifuge

Date: 07/28/2014

PCR of GAFP1+B0015 = K1364007 (pSB1C3) A, B, C, D, E, F + electrophoresis

Date: 07/28/2014
Result: clones B, C, D, E, F seem to have the expected construction.

Digestion of clones E, F of GAFP1+Ter B0015 = K1364007 with EcoRI and PstI + electrophoresis

Date: 07/28/2014
Result: clones B, C, D, E, F have the expected construction.

4. Cloning GAFP1+terminator with Pveg on pSB1C3 (K1364008) in E. coli

Digestion of GAFP1+ter = K1364007 on pSB1C3 and K823003 (terminator on pSB1C3) + electrophoresis

Date: 07/29/2014

Gel extraction of K1364007 (extraction of GAFP1+ter gene)

Date: 07/29/2014

Ligation of GAFP1+ter in K823003 (Pveg on pSB1C3)

Date: 07/29/2014
Result: 20µL ligation of GAFP1+ter in K823003

Transformation of ligation products in E.coli

Date: 07/29/2014

Culture of 8 clones: A, B, C, D, E, F, G, H of transformed E. coli

Date: 07/30/2014

QIAprep Spin Miniprep Kit Using a Microcentrifuge

Date: 07/31/2014

PCR of GAFP1+B0015 + K823003 = K1364008 (pSB1C3) A, B, C, D, E, F, G, H + electrophoresis

Date: 31/07/2014
Result: clones A, B, C, D, E, G, H seem to have the expected construction

Digestion of clones A, B, C, D, E, G, H of Pveg+K1364007 = K1364008 with EcoRI and PstI + electrophoresis

Date: 31/07/2014
Result: clones A, B, C, D, E, G, H have the expected construction

5. Cloning Pveg+GAFP1+Ter B0015 (BBa_K1364008) on pSBBS4S (BBa_K823022)

Digestion of Pveg+GAFP1+ ter B0015 (BBa_K1364008) on pSB1C3 and PsBBs4S (BBa_K823022)

Date: 08/01/2014

Ligation of K134008 (Pveg+GAFP1+ter fragment) and K823022 (PsBBs4S)

Date: 08/01/2014

Transformation of ligation products in E.coli

Date: 08/01/2014

Culture of 8 clones: A, B, C, D, E, F, G, H of transformed E. coli

Date: 08/02/2014

QIAprep Spin Miniprep Kit Using a Microcentrifuge

Date: 08/04/2014

Digestion of clones A, B, C, D, E, G, H of K1364008+K823022 with EcoRI and PstI + electrophoresis

Date: 08/04/2014
Result: clones A, E, F have the right construction

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EcAMP

The EcAMP part was sent by the Utah iGEM team. It was on a pUC plasmid (ampicilin resistance) with BioBrick suffix and prefix.

1. Transformation of EcAMP in Escherichia coli MC 1061

Date: 07/25/2014

2. Spreading of coli cells transformed with EcAMP (plasmid pUC)

Date: 07/28/2014

3. Liquid culture, Miniprep and Test of the miniprep

Date: 07/30/2014

4. Cloning 1: EcAMP + P_veg + RBS

Digestion of EcAMP (insert) by XbaI and PstI

Date: 07/31/2014

Digestion of P^veg + RBS (VECTOR)

Date: 07/31/2014

Ligation and transformation

Date: 08/04/2014

PCR test

Date: 08/05/2014

Analytical digestion

Date: 08/05/2014
Result: The first cloning show a lack of DNA in the Miniprep EcAMP. The bands are hardly visible on the gel for the linearization of EcAMP + Pveg + RBS. The problem came from the PCR cleanup step: the fragment size of EcAMP is lower than 200bp.

5. Cloning 2: EcAMP + Pveg + RBS

Date: 08/06/2014

Digestion of EcAMP (insert) by XbaI and PstI

Date: 08/06/2014

Digestion of P_veg + RBS (vector)

Date: 08/06/2014

Heat inactivation of the enzymes

Date: 08/06/2014

Ligation and transformation

Date: 08/06/2014

PCR test

Date: 08/07/2014

Striation on a petri dish to purify the clone

Purpose: to isolate a clone with vector+insert
Date: 08/072014

Miniprep of P_veg + SpoVG + EcAMP and analytic digestion

Date: 08/08/2014

Ligation of Pveg SpoVG EcAMP with double terminateur B0015 + transformation and liquid culture

Date: 08/11/2014

Miniprep of P_veg SpoVG EcAMP + analytic digestion

Date: 08/13/2014

Cloning K1364011 (EcAMP + P_veg +SpoVG + B0015) + K823022 (pSB_BS4S): Digestion, ligation, transformation

Date: 08/19/2014

Cloning K1364011 + K823023 (pSB_BS1C) : Digestion, ligation, transformation

Date: 08/18/2014

Verification of the insertion of K1364011 + K823022 (pSB_BS4S) into the subtilis genome by threonine test

Date: 08/21/2014

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Assembling the fungicides module: D4E1 + GAFP1

1. Cloning GAFP1+D4E1 on pSB1C3: BBa_K1364012

Digestion of D4E1 on pEX-A2 and GAFP1 on pSB1C3

Date: 07/28/2014
Result: We obtained 40µL digestion of D4E1 on pEX-A2 and of GAFP1 on pSB1C3.

Ligation of digestions of D4E1 on pEX-A2 and of GAFP1 on pSB1C3

Date: 07/28/2014

Transformation of ligation of GAFP1 and D4E1 on pSB1C3 into E. coli

Date: 07/28/2014

PCR of GAFP1 + D4E1 (pSB1C3) A, B, C, D, E, F + electrophoresis

Date: 07/29/2014
Result: clones A, C, F, G seem to have the expected construction.

Digestion of GAFP1 + D4E1 (pSB1C3) A, C, F, G + electrophoresis

Date: 07/30/2014
Result: clone F (BBa_K1364012) has the expected construction and is placed on cryopreservation.

2. Cloning GAFP1 + D4E1 (BBa_K1364012) on P_veg plasmid (BBa_K823003): BBa_K1364013

Date: 08/04/2014

3. Cloning P_veg + GAFP1 + D4E1 (BBa_K1364013) on pSB_{BS_{4S (BBa_K823022)}}

Date: 08/06/2014

4. Cloning P_veg + GAFP1 + D4E1 (BBa_K1364013) on pSB_BS1C lacZ (BBa_K823023)

Date: 08/11/2014

5. Fungicides tests

Date: 08/15/2014

Cloning D4E1-GAFP1-EcAMP: BBa_K1364014

1. Construction of BBa_K1364014 in pSB1C3, in E. coli

Digestion of BBa_K1364010 and BBa_K1364012

Digestion of BBa_K1364010 by SpeI and PstI, and digestion of BBa_K1364012 by XbaI and PstI.
Date: 08/11/2014
Result: We obtained digested fragments of BBa_K1364012 and ~500 bp fragment of BBa_K1364010

Ligation of ~500 bp fragment from BBa_K1364010 and BBa_K1364012

Date: 08/11/2014
Result: We obtained ligation of K1364012 and ~500 bp fragment of BBa_K1364010

Transformation of ligation of ~500 bp fragment from BBa_K1364010 and BBa_K1364012 = BBa_K1364014 in E.coli

Date: 08/12/2014

Testing 8 E.coli + BBa_K1364014 clones

Date: 08/14/2014

Testing 15 E.coli + BBa_K1364014 clones

Date: 08/18/2014
Result: clones H, J, O, Q, U, V might have the right BBa_K1364014 construction

2. BBa_K1364014 in pSB_BS4S (BBa_K823022) and pSB_BS1C (BBa_K823023)

Digestion of BBa_K1364014, BBa_K823022 and BBa_K823023

Date: 08/22/2014
Result: Digestion of BBa_K1364014, BBa_K823022 and BBa_K823023 by EcoRI and PstI were well performed.

Ligation of BBa_K1364014 with BBa_K823022 and K1364014 with K823023

Date: 08/22/2014
Result: We obtained 20µL ligation of K1364014 with BBa_K823022 and of BBa_K1364014 with BBa_K823023.

Transformation of ligations in E. coli

E. coli transformed by BBa_K1364014 + BBa_K823022 spread on LB + Amp 100µg/mL agar plate

E. coli transformed by BBa_K1364014 + BBa_K823023 spread on LB + Amp 100µg/mL agar plate
Date: 08/25/2014

Testing E. coli + BBa_K1364014 + BBa_K823022 and E. coli + BBa_K1364014 + BBa_K823023 clones

Date: 08/27/2014

3. Transformation of BBa_K1364014+BBa_K823022 and BBa_K1364014 + BBa_K823023 in B. subtilis

B. subtilis transformed by 10µL BBa_K1364014+BBa_K823022 spread on LB+Spec 75µg/mL agar plate
B. subtilis transformed by 10µL BBa_K1364014+BBa_K823023 spread on LB+Cm 15µg/mL agar plate
Date: 08/28/2014

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Fungicides tests

1. D4E1-GAFP1

Transformation of P_veg - D4E1 - GAFP1 in Bacillus subtilis on pSB_BS4S

Date: 08/12/2014

Integration threonine test + fungicide test

Date: 08/13/2014

Cloning D4E1 into pSB_BS1C + fungicide test

Date:08/15/2014

D4E1 on pSB1C3 + fungicide test

Date:08/19/2014

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@@ Line 333: / Line 333: @@
 <div class="thelanguage">
-<p class="title2">D4E1</p>
+<div class="technology2">D4E1</div>
+<div class="thelanguage2">
 <p class="title2">1.	Amplification of synthetic gene (D4E1 on pEX-A2)</p>
@@ Line 402: / Line 403: @@
 <br><b>Result:</b> clones C, D have the expected construction and are placed in cryopreservation.</p>
-<p class="title2">4. Cloning Pveg+D4E1 on pSBBS4S (K823022) </p>
+<p class="title2">4. Cloning P<sub>veg</sub> + D4E1 on pSB<sub>BS</sub>4S (K823022) </p>
 <p class="texte"><b>Date:</b> 08/13/2014</p>
-<p class="title2">5. Cloning Pveg + D4E1 on pSBBS1C lacZ (23)</p>
+<p class="title2">5. Cloning P<sub>veg</sub> + D4E1 on pSB<sub>BS</sub>1C lacZ (23)</p>
+<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 2); return false">Collapse</a></p>
+</div>
+<div class="technology2">GAFP1</div>
+<div class="thelanguage2">
-<p class="title2">GAFP1</p>
 <p class="title2">1. Amplification of synthetic gene (GAFP1 on pEX-A2)</p>
@@ Line 533: / Line 539: @@
 <br><b>Result:</b> clones A, E, F have the right construction</p>
+<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 2); return false">Collapse</a></p>
+</div>
+<div class="technology2">EcAMP</div>
+<div class="thelanguage2">
+<p class="texte">The EcAMP part was sent by the Utah iGEM team. It was on a pUC plasmid (ampicilin resistance) with BioBrick suffix and prefix.</p>
-<p class="title2">EcAMP</p>
+<p class="title2">1.	Transformation of EcAMP in <i>Escherichia coli</i> MC 1061</p>
-<p class="texte">The EcAMP part was sent by the Utah iGEM team. It was on a pUC plasmid (ampicilin resistance) with biobrick suffix and prefix.</p>
-<p class="title2">1.	Transformation of EcAMP in Escherichia coli MC 1061</p>
 <p class="texte"><b>Date:</b> 07/25/2014</p>
-<p class="title2">2.	Spreading of coli cells transformed with pUC + Utah </p>
+<p class="title2">2.	Spreading of coli cells transformed with EcAMP (plasmid pUC) </p>
 <p class="texte"><b>Date:</b> 07/28/2014</p>
-<p class="title2">3.	Liquid culture + Miniprep + Test of the miniprep</p>
+<p class="title2">3.	Liquid culture, Miniprep and Test of the miniprep</p>
 <p class="texte"><b>Date:</b> 07/30/2014</p>
-<p class="title2">4.	Cloning 1: EcAMP + Pveg + RBS</p>
+<p class="title2">4.	Cloning 1: EcAMP + P<sub>veg</sub> + RBS</p>
-<p class="title3">Digestion of EcAMP (INSERT) by XbaI and PstI</p>
+<p class="title3">Digestion of EcAMP (insert) by XbaI and PstI</p>
 <p class="texte"><b>Date:</b> 07/31/2014</p>
-<p class="title3">Digestion of Pveg + RBS (VECTOR)</p>
+<p class="title3">Digestion of P<sup>veg</sup> + RBS (VECTOR)</p>
 <p class="texte"><b>Date:</b> 07/31/2014</p>
@@ Line 568: / Line 575: @@
 <p class="texte"><b>Date:</b> 08/05/2014
-<br>The first cloning show a lack of DNA in the Miniprep EcAMP. The bands are hardly visible on the gel for the linearization of EcAMP + Pveg + RBS. The problem came from the PCR cleanup step: the fragment size of EcAMP is lower than 200bp.</p>
+<br><b>Result:</b> The first cloning show a lack of DNA in the Miniprep EcAMP. The bands are hardly visible on the gel for the linearization of EcAMP + Pveg + RBS. The problem came from the PCR cleanup step: the fragment size of EcAMP is lower than 200bp.</p>
 <p class="title2">5.	 Cloning 2: EcAMP + Pveg + RBS </p>
-<p class="texte"><b>Date:</b> 06/08/2014</p>
+<p class="texte"><b>Date:</b> 08/06/2014</p>
-<p class="title3">Digestion of EcAMP (INSERT) by XbaI and PstI</p>
+<p class="title3">Digestion of EcAMP (insert) by XbaI and PstI</p>
-<p class="title3">Digestion of Pveg + RBS (VECTOR)</p>
+<p class="texte"><b>Date:</b> 08/06/2014</p>
+<p class="title3">Digestion of P<sub>veg</sub> + RBS (vector)</p>
+<p class="texte"><b>Date:</b> 08/06/2014</p>
 <p class="title3">Heat inactivation of the enzymes</p>
+<p class="texte"><b>Date:</b> 08/06/2014</p>
 <p class="title3">Ligation and transformation</p>
+<p class="texte"><b>Date:</b> 08/06/2014</p>
 <p class="title3">PCR test</p>
-<p class="texte"><b>Date:</b> 07/08/2014</p>
+<p class="texte"><b>Date:</b> 08/07/2014</p>
 <p class="title3">Striation on a petri dish to purify the clone</p>
-<p class="texte">Purpose: to isolate a clone with vector+insert
+<p class="texte"><b>Purpose:</b> to isolate a clone with vector+insert
 <br><b>Date:</b> 08/072014</p>
-<p class="title3">Miniprep of Pveg+SpoVG+EcAMP and analytic digestion</p>
+<p class="title3">Miniprep of P<sub>veg</sub> + SpoVG + EcAMP and analytic digestion</p>
 <p class="texte"><b>Date:</b> 08/08/2014</p>
-<p class="title3">Ligation of Pveg SpoVG EcAMP with double terminateur B0015 +Transformation and liquid culture</p>
+<p class="title3">Ligation of Pveg SpoVG EcAMP with double terminateur B0015 + transformation and liquid culture</p>
 <p class="texte"><b>Date:</b> 08/11/2014</p>
-<p class="title3">Miniprep of Pveg SpoVG EcAMP + analytic digestion</p>
+<p class="title3">Miniprep of P<sub>veg</sub> SpoVG EcAMP + analytic digestion</p>
 <p class="texte"><b>Date:</b> 08/13/2014</p>
-<p class="title3">Cloning K1364011 (EcAMP+Pveg +SpoVG + B0015) + K823022 (psBbs4S) : Digestion, ligation, transformation</p>
+<p class="title3">Cloning K1364011 (EcAMP + P<sub>veg</sub> +SpoVG + B0015) + K823022 (pSB<sub>BS</sub>4S): Digestion, ligation, transformation</p>
 <p class="texte"><b>Date:</b> 08/19/2014</p>
-<p class="title3">Cloning K1364011 + K823023 (pSBBS1C) : Digestion, ligation, transformation</p>
+<p class="title3">Cloning K1364011 + K823023 (pSB<sub>BS</sub>1C) : Digestion, ligation, transformation</p>
 <p class="texte"><b>Date:</b> 08/18/2014</p>
-<p class="title3">Verification of the insertion of K1364011 + K823022 (pSBBS4S) into the subtilis genome by threonine test </p>
+<p class="title3">Verification of the insertion of K1364011 + K823022 (pSB<sub>BS</sub>4S) into the subtilis genome by threonine test </p>
-<p class="texte"><b>Date:</b> 08/21/2014
+<p class="texte"><b>Date:</b> 08/21/2014</p>
-<br>
-<br>Assembling the fungicides module : </p>
+<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 2); return false">Collapse</a></p>
+</div>
+<div class="technology2">Assembling the fungicides module: D4E1 + GAFP1</div>
+<div class="thelanguage2">
-<p class="title2">D4E1 + GAFP1 </p>
 <p class="title2">1.  Cloning GAFP1+D4E1 on pSB1C3: <a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a></p>
@@ Line 622: / Line 633: @@
 <p class="texte"><b>Date:</b> 07/28/2014</p>
-<p class="title3">PCR of GAFP1+D4E1 (pSB1C3) A, B, C, D, E, F + electrophoresis</p>
+<p class="title3">PCR of GAFP1 + D4E1 (pSB1C3) A, B, C, D, E, F + electrophoresis</p>
 <p class="texte"><b>Date:</b> 07/29/2014
 <br><b>Result:</b> clones A, C, F, G seem to have the expected construction.</p>
-<p class="title3">Digestion of GAFP1+D4E1 (pSB1C3) A, C, F, G + electrophoresis</p>
+<p class="title3">Digestion of GAFP1 + D4E1 (pSB1C3) A, C, F, G + electrophoresis</p>
 <p class="texte"><b>Date:</b> 07/30/2014
 <br><b>Result:</b> clone  F (<a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a>) has the expected construction and is placed on cryopreservation.</p>
@@ Line 632: / Line 643: @@
-<p class="title2">2.	 Cloning GAFP1+D4E1 (<a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a>) on Pveg plasmid (<a href="http://parts.igem.org/Part:BBa_K823003">BBa_K823003</a>): <a href="http://parts.igem.org/Part:BBa_K1364013">BBa_K1364013</a></p>
+<p class="title2">2.	 Cloning GAFP1 + D4E1 (<a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a>) on P<sub>veg</sub> plasmid (<a href="http://parts.igem.org/Part:BBa_K823003">BBa_K823003</a>): <a href="http://parts.igem.org/Part:BBa_K1364013">BBa_K1364013</a></p>
-<p class="title2">3.	 Cloning Pveg+GAFP1+D4E1 (<a href="http://parts.igem.org/Part:BBa_K1364013">BBa_K1364013</a>) on pSBBS4S (<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a>)</p>
+<p class="texte"><b>Date:</b> 08/04/2014
-<p class="title2">4.	 Cloning Pveg+GAFP1+D4E1 (<a href="http://parts.igem.org/Part:BBa_K1364013">BBa_K1364013</a>) on pSBBS1C lacZ (<a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a>)</p>
-<p class="title2">5.	 Fungicides tests</p>
+<p class="title2">3.	 Cloning P<sub>veg</sub> + GAFP1 + D4E1 (<a href="http://parts.igem.org/Part:BBa_K1364013">BBa_K1364013</a>) on pSB<sub>BS<sub>4S (<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a>)</p>
+<p class="texte"><b>Date:</b> 08/06/2014
+<p class="title2">4.	 Cloning P<sub>veg</sub> + GAFP1 + D4E1 (<a href="http://parts.igem.org/Part:BBa_K1364013">BBa_K1364013</a>) on pSB<sub>BS</sub>1C lacZ (<a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a>)</p>
+<p class="texte"><b>Date:</b> 08/11/2014
+<p class="title2">5.	 Fungicides tests</p>
+<p class="texte"><b>Date:</b> 08/15/2014
 <p class="title2">Cloning D4E1-GAFP1-EcAMP: <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a></p>
-<p class="title2">1.  Construction of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> in PsB1C3, in E. coli</p>
+<p class="title2">1.  Construction of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> in pSB1C3, in <i>E. coli</i></p>
 <p class="title3">Digestion of <a href="http://parts.igem.org/Part:BBa_K1364010">BBa_K1364010</a> and <a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a></p>
-<p class="texte">Digestion of <a href="http://parts.igem.org/Part:BBa_K1364010">BBa_K1364010</a> by Spe1 and Pst1, and digestion of <a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a> by XbaI and Pst1.
+<p class="texte">Digestion of <a href="http://parts.igem.org/Part:BBa_K1364010">BBa_K1364010</a> by SpeI and PstI, and digestion of <a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a> by XbaI and PstI.
 <br><b>Date:</b> 08/11/2014
 <br><b>Result:</b> We obtained digested fragments of <a href="http://parts.igem.org/Part:BBa_K1364012">BBa_K1364012</a> and ~500 bp fragment of <a href="http://parts.igem.org/Part:BBa_K1364010">BBa_K1364010</a></p>
@@ Line 663: / Line 680: @@
-<p class="title2">2. <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> in PsBs4s (<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a>) and PsBs1C (<a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a>)</p>
+<p class="title2">2. <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> in pSB<sub>BS</sub>4S (<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a>) and pSB<sub>BS</sub>1C (<a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a>)</p>
 <p class="title3">Digestion of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>, <a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and <a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a></p>
 <p class="texte"><b>Date:</b> 08/22/2014
-<br><b>Result</b>: digestion of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>, <a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and <a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a> by EcoR1 and Pst1 were well performed.</p>
+<br><b>Result</b>: Digestion of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>, <a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and <a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a> by EcoRI and PstI were well performed.</p>
 <p class="title3">Ligation of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> with <a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and K1364014 with K823023</p>
@@ Line 673: / Line 690: @@
 </p>
 <p class="title3">Transformation of ligations in <i>E. coli</i></p>
-<p class="texte">E. coli transformed by <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>+<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> spread on LB+Amp 100µg/mL agar plate</p>
+<p class="texte"><I>E. coli</I> transformed by <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> + <a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> spread on LB + Amp 100µg/mL agar plate</p>
-<p class="texte">E. coli transformed by <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>+<a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a> spread on LB+Amp 100µg/mL agar plate
+<p class="texte"><i>E. coli</i> transformed by <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> + <a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a> spread on LB + Amp 100µg/mL agar plate
 <br><b>Date:</b> 08/25/2014</p>
@@ Line 682: / Line 699: @@
-<p class="title2">3.  Transformation of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>+<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>+<a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a> in <i>B. subtilis</i></p>
+<p class="title2">3.  Transformation of <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>+<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> and <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a> + <a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a> in <i>B. subtilis</i></p>
 <p class="texte"><i>B. subtilis</i> transformed by 10µL <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>+<a href="http://parts.igem.org/Part:BBa_K823022">BBa_K823022</a> spread on LB+Spec 75µg/mL agar plate
 <br><i>B. subtilis</i> transformed by 10µL <a href="http://parts.igem.org/Part:BBa_K1364014">BBa_K1364014</a>+<a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a>  spread on LB+Cm 15µg/mL agar plate
 <br><b>Date:</b> 08/28/2014</p>
-<p class="title2">Fungicides tests</p>
+<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 2); return false">Collapse</a></p>
+</div>
+<div class="technology2">Fungicides tests</div>
+<div class="thelanguage2">
 <p class="title2">1.	D4E1-GAFP1</p>
-<p class="title3">Transformation in bacillus  Pveg-D4E1-GAFP1 on pSBBS4S </p>
+<p class="title3">Transformation of P<sub>veg</sub> - D4E1 - GAFP1 in <i>Bacillus subtilis</i>  on pSB<sub>BS</sub>4S </p>
 <p class="texte"><b>Date:</b> 08/12/2014</p>
 <p class="title3">Integration threonine test + fungicide test </p>
@@ Line 696: / Line 718: @@
 <p class="title2"2.	D4E1</p>
 <p class="title3">
-Cloning D4E1 into pSBBS1C + fungicide test</p>
+Cloning D4E1 into pSB<sub>BS</sub>1C + fungicide test</p>
 <p class="texte"><b>Date:</b>08/15/2014</p>
 <p class="title3">
@@ Line 707: / Line 729: @@
 <div class="clear"></div>
+</div>
      </div>
    </div>

Team:Toulouse/Notebook/project-monitoring

From 2014.igem.org

Latest revision as of 01:22, 18 October 2014

iGEM Toulouse 2014

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