Difference between revisions of "Team:UNC-Chapel Hill/Notebook"

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<h4 style = "font-size: 24 px"> May 18th, 2015 </h>
 
<h4 style = "font-size: 24 px"> May 18th, 2015 </h>
−
    <p>
+
<p>
−
        Connor and Sean
+
Connor and Sean
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Resuspended parts BBa_E1010 and BBa_I716120 on plate 3 well 11n and plate 3 well 11m, respectively.  Transformed both and placed in 37 degree incubator
 
Resuspended parts BBa_E1010 and BBa_I716120 on plate 3 well 11n and plate 3 well 11m, respectively.  Transformed both and placed in 37 degree incubator
 
*Note: parts left in 37 degree incubator too long (>24 hours) and overgrew
 
*Note: parts left in 37 degree incubator too long (>24 hours) and overgrew
Line 69: Line 71:
 
<p>
 
<p>
 
May 21st, 2015
 
May 21st, 2015
 +
<p>
 
Elliot and Danny
 
Elliot and Danny
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Resuspended part BBa_K118011 on plate 3 well 20c. Transformed this part along with BBa_E1010 again.
 
Resuspended part BBa_K118011 on plate 3 well 20c. Transformed this part along with BBa_E1010 again.
 
</p>
 
</p>
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<p>
 
<p>
 
Connor and Sean
 
Connor and Sean
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Resuspended parts BBa_K1033916 and BBa_K592009 on plate 4 well 6m and plate 1 well 15e, respectively. Transformed those parts and BBa_E1010.
 
Resuspended parts BBa_K1033916 and BBa_K592009 on plate 4 well 6m and plate 1 well 15e, respectively. Transformed those parts and BBa_E1010.
 +
<p>
 
*Note: BBa_K1033916 plate was thrown out due to suspicion of LBKanR mislabeling and the two other chromoproteins appear to have been switched.
 
*Note: BBa_K1033916 plate was thrown out due to suspicion of LBKanR mislabeling and the two other chromoproteins appear to have been switched.
 +
<p>
 
Elliot and Danny
 
Elliot and Danny
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Transformed parts BBa_E1010, BBa_K1033916, BBa_K592009,  BBa_K861171 and BBa_K118011.
 
Transformed parts BBa_E1010, BBa_K1033916, BBa_K592009,  BBa_K861171 and BBa_K118011.
 +
<p>
 
Will and Jerry
 
Will and Jerry
 +
<p>
 
ZR Plasmid Miniprep Classic
 
ZR Plasmid Miniprep Classic
 +
<p>
 
Miniprepped parts BBa_K861171 and BBa_K118011. Prepared 1x TAE Buffer from stock.
 
Miniprepped parts BBa_K861171 and BBa_K118011. Prepared 1x TAE Buffer from stock.
 
</p>
 
</p>
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<p>
 
<p>
 
Will
 
Will
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Did overall inventory and plated BBa_K861171
 
Did overall inventory and plated BBa_K861171
 +
<p>
 
Connor and Sean
 
Connor and Sean
 +
<p>
 
Liquid Cultures
 
Liquid Cultures
−
Created liquid cultures of BBa_E1010 (2), BBa_K592009 (2), and BBa_K1033916 and placed in 37 degree shaking incubator overnight
+
<p>
 +
Created liquid cultures of BBa_E1010 (2), BBa_K592009 (2), and BBa_K1033916 and placed in 37 degree shaking incubator overnight.
 +
<p>
 
*Note: Liquid cultures spent >24 hours in 37 degree incubator, so thrown out. Also, BBa_K118011 failed to grow.
 
*Note: Liquid cultures spent >24 hours in 37 degree incubator, so thrown out. Also, BBa_K118011 failed to grow.
 
</p>
 
</p>
Line 99: Line 119:
 
<p>
 
<p>
 
Elliot and Danny
 
Elliot and Danny
 +
<p>
 
Liquid Cultures
 
Liquid Cultures
 +
<p>
 
Created liquid cultures of BBa_E1010 (2), BBa_K592009 (2), and BBa_K1033916 and placed in 37 degree shaking incubator overnight. LB was autoclaved and placed in 4 degree fridge.  
 
Created liquid cultures of BBa_E1010 (2), BBa_K592009 (2), and BBa_K1033916 and placed in 37 degree shaking incubator overnight. LB was autoclaved and placed in 4 degree fridge.  
 
</p>
 
</p>
Line 105: Line 127:
 
<p>
 
<p>
 
Elliot and Danny
 
Elliot and Danny
 +
<p>
 
ZR Plasmid Miniprep Classic
 
ZR Plasmid Miniprep Classic
 +
<p>
 
Miniprepped BBa_E1010 (2), BBa_K592009 (2), and BBa_K1033916 and nanodropped for concentrations.  
 
Miniprepped BBa_E1010 (2), BBa_K592009 (2), and BBa_K1033916 and nanodropped for concentrations.  
 
</p>
 
</p>
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<p>
 
<p>
 
Will
 
Will
 +
<p>
 
Did overall inventory and made a 1L DI water stock
 
Did overall inventory and made a 1L DI water stock
 
</p>
 
</p>
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<p>
 
<p>
 
Will and Jerry
 
Will and Jerry
 +
<p>
 
PCR
 
PCR
 +
<p>
 
Received and reconstituted MLC Parts and MLC primers. Performed PCR amplification on the MLC parts.
 
Received and reconstituted MLC Parts and MLC primers. Performed PCR amplification on the MLC parts.
 
</p>
 
</p>
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<p>
 
<p>
 
Will and Jerry
 
Will and Jerry
 +
<p>
 
Agarose Gel Electrophoresis, DNA Clean + Concentrator Prep, PCR
 
Agarose Gel Electrophoresis, DNA Clean + Concentrator Prep, PCR
−
Ran a gel verifying the MLC parts, concentrated the PCR-generated plasmids and nanodropped. After determining that concentrations were insufficient, ran another PCR for MLC amplification. Also conducted inventory.  
+
<p>
 +
Ran a gel verifying the MLC parts, concentrated the PCR-generated plasmids and nanodropped. After determining that concentrations were insufficient, ran another PCR for MLC amplification. Also conducted inventory.
 +
<p>
 
Connor and Sean
 
Connor and Sean
 +
<p>
 
DNA Clean + Concentrator Prep, Transformation
 
DNA Clean + Concentrator Prep, Transformation
−
Prepped the DNA and transformed BBa_J4450, which was reconstituted from plate 4 well 4b. Made a glycerol stock  
+
<p>
 +
Prepped the DNA and transformed BBa_J4450, which was reconstituted from plate 4 well 4b. Made a glycerol stock.
 +
<p>
 
Elliot and Danny
 
Elliot and Danny
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Created ampicillin plates and reconstituted BBa_J61100 on plate 4 well b4. Transformed BBa_J61100 on the ampicillin plates.
 
Created ampicillin plates and reconstituted BBa_J61100 on plate 4 well b4. Transformed BBa_J61100 on the ampicillin plates.
 
</p>
 
</p>
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<p>
 
<p>
 
Connor, Sean and Elliot
 
Connor, Sean and Elliot
 +
<p>
 
Liquid Cultures, DNA Clean and Concentrator Prep
 
Liquid Cultures, DNA Clean and Concentrator Prep
 +
<p>
 
Made 2 liquid cultures of BBa_J61100 (LBAmp) and 4 liquid cultures of BBa_J4450 (LBCam). Prepped the new MLC PCR mixes and nanodropped for concentrations.
 
Made 2 liquid cultures of BBa_J61100 (LBAmp) and 4 liquid cultures of BBa_J4450 (LBCam). Prepped the new MLC PCR mixes and nanodropped for concentrations.
 
</p>
 
</p>
Line 140: Line 177:
 
<p>
 
<p>
 
Elliot and Danny
 
Elliot and Danny
 +
<p>
 
ZR Plasmid Miniprep Classic
 
ZR Plasmid Miniprep Classic
 +
<p>
 
Miniprepped the liquid cultures of BBa_J61100 and BBa_J4450 (6 in total). Stored in the -20 degrees freezer.
 
Miniprepped the liquid cultures of BBa_J61100 and BBa_J4450 (6 in total). Stored in the -20 degrees freezer.
 
</p>
 
</p>
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<p>
 
<p>
 
Elliot and Danny
 
Elliot and Danny
 +
<p>
 
Liquid Cultures
 
Liquid Cultures
 +
<p>
 
Created 3 liquid cultures of BBa_J4450 (LBCam) and 6 liquid cultures of BBa_J61100 (LBAmp).
 
Created 3 liquid cultures of BBa_J4450 (LBCam) and 6 liquid cultures of BBa_J61100 (LBAmp).
 
</p>
 
</p>
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<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
ZR Plasmid Miniprep Classic
 
ZR Plasmid Miniprep Classic
 +
<p>
 
Miniprepped 2 BBa_J4450 cultures and 5 BBa_J61100 cultures. Nanodropped and stored in the -20 degrees freezer.
 
Miniprepped 2 BBa_J4450 cultures and 5 BBa_J61100 cultures. Nanodropped and stored in the -20 degrees freezer.
 +
<p>
 
Elliot and Danny
 
Elliot and Danny
 +
<p>
 
3A Assembly, Agarose Gel Electrophoresis
 
3A Assembly, Agarose Gel Electrophoresis
 +
<p>
 
Conducted 3A Assembly with the following components: pSB1C3 (from BBa_J4450), BBa_K1033916/BBa_E1010/BBa_K592009 (downstream component) and BBa_J61100 (upstream component). Made a 1% agarose gel with the following wells: 1: 1 kb ladder; 2: BBa_E1010 #3; 3. BBa_E1010 #2; 4. BBa_K1033916 #3; 5. BBa_K592009 #2; 6. BBa_K861171 # 3; 7. 1 kb ladder. Made another 1% agarose gel: 1: 1 kb ladder; 2: 100 bp ladder; 3. BBa_K1033916 #3; 4. BBa_K592009 #2; 5. BBa_K1033916 #1; 6. BBa_K861171 #1; 7. BBa_K861171 #2; 8. BBa_K861171 #3
 
Conducted 3A Assembly with the following components: pSB1C3 (from BBa_J4450), BBa_K1033916/BBa_E1010/BBa_K592009 (downstream component) and BBa_J61100 (upstream component). Made a 1% agarose gel with the following wells: 1: 1 kb ladder; 2: BBa_E1010 #3; 3. BBa_E1010 #2; 4. BBa_K1033916 #3; 5. BBa_K592009 #2; 6. BBa_K861171 # 3; 7. 1 kb ladder. Made another 1% agarose gel: 1: 1 kb ladder; 2: 100 bp ladder; 3. BBa_K1033916 #3; 4. BBa_K592009 #2; 5. BBa_K1033916 #1; 6. BBa_K861171 #1; 7. BBa_K861171 #2; 8. BBa_K861171 #3
 
</p>
 
</p>
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<p>
 
<p>
 
Elliot + Danny + Jerry
 
Elliot + Danny + Jerry
 +
<p>
 
Agarose Gel Electrophoresis
 
Agarose Gel Electrophoresis
 +
<p>
 
Ran a diagnostic 2% gel for the repressible.  
 
Ran a diagnostic 2% gel for the repressible.  
 +
<p>
 
Will + Jerry
 
Will + Jerry
 +
<p>
 
Agarose Gel Electrophoresis
 
Agarose Gel Electrophoresis
 +
<p>
 
Ran a 1% gel with the following specifications: 1: 1 kb ladder; 2: 100 bp ladder; 3: BBa_K1033916 #1; 4: BBa_K1033916 #2; 5: BBa_K1033916 #3; 6: BBa_K592009 #2; 7: BBa_K861171 #1; 8: BBa_K861171 #2; 9: BBa_K861171 #3
 
Ran a 1% gel with the following specifications: 1: 1 kb ladder; 2: 100 bp ladder; 3: BBa_K1033916 #1; 4: BBa_K1033916 #2; 5: BBa_K1033916 #3; 6: BBa_K592009 #2; 7: BBa_K861171 #1; 8: BBa_K861171 #2; 9: BBa_K861171 #3
 +
<p>
 
Connor + Will
 
Connor + Will
 +
<p>
 
3A Assembly, Simple Recombination
 
3A Assembly, Simple Recombination
 +
<p>
 
Conducted a 3A assembly with the following components: pSB1C3, BBa_E1010/BBa_K592009, BBa_J1100. Conducted simple recombination (cutting a backbone and “part” with the same enzymes) with the following components: MLC1/2/3/4, pSB1C3.
 
Conducted a 3A assembly with the following components: pSB1C3, BBa_E1010/BBa_K592009, BBa_J1100. Conducted simple recombination (cutting a backbone and “part” with the same enzymes) with the following components: MLC1/2/3/4, pSB1C3.
 +
<p>
 
Will
 
Will
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Transformed the two 3A plasmids and four simple recombination plasmids.
 
Transformed the two 3A plasmids and four simple recombination plasmids.
 +
<p>
 
*Note: all MLC plates failed to grow
 
*Note: all MLC plates failed to grow
 
</p>
 
</p>
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<p>
 
<p>
 
Will
 
Will
 +
<p>
 
PCR, Transformation
 
PCR, Transformation
 +
<p>
 
Conducted another PCR amplification procedure for the order MLC parts (1-4). Transformed the MLC + pSB1C3 constructs that failed to grow once more.
 
Conducted another PCR amplification procedure for the order MLC parts (1-4). Transformed the MLC + pSB1C3 constructs that failed to grow once more.
 
</p>
 
</p>
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<p>
 
<p>
 
Will
 
Will
 +
<p>
 
Agarose Gel Electrophoresis, Gel DNA Recovery Prep
 
Agarose Gel Electrophoresis, Gel DNA Recovery Prep
 +
<p>
 
Made glycerol stocks of BBa_K1033916#3/pSB1C3/BBa_J61100 (pink, so actually BBa_J4450), BBa_K592009#2/pSB1C3/BBa_J61100, BBa_E1010#2/pSB1C3/BBa_J61100. Then ran a 1% gel with the following specifications: 1: 1 kb ladder; 2: 100 bp ladder; 3: MLC1: 4: MLC2; 5: MLC3; 6: MLC4; 7: MasterMix; 8: USP H20. Then ran a DNA gel recovery prep and nanodropped.
 
Made glycerol stocks of BBa_K1033916#3/pSB1C3/BBa_J61100 (pink, so actually BBa_J4450), BBa_K592009#2/pSB1C3/BBa_J61100, BBa_E1010#2/pSB1C3/BBa_J61100. Then ran a 1% gel with the following specifications: 1: 1 kb ladder; 2: 100 bp ladder; 3: MLC1: 4: MLC2; 5: MLC3; 6: MLC4; 7: MasterMix; 8: USP H20. Then ran a DNA gel recovery prep and nanodropped.
−
*Note: MLC remainders from the gel were not enough, so another PCR is recommended
+
*Note: MLC remainders from the gel were not enough, so another PCR is recommended.
 +
<p>
 
Connor
 
Connor
 +
<p>
 
Zyppy Plasmid Miniprep, Transformation
 
Zyppy Plasmid Miniprep, Transformation
 +
<p>
 
Miniprepped BBa_K592009/BBa_J61100/pSB1C3 and BBa_E1010/BBa_J61100/pSB1C3 liquid cultures. Nanodropped the results. Transformed the BBa_1033916 #1/#2 3A results.  
 
Miniprepped BBa_K592009/BBa_J61100/pSB1C3 and BBa_E1010/BBa_J61100/pSB1C3 liquid cultures. Nanodropped the results. Transformed the BBa_1033916 #1/#2 3A results.  
 +
<p>
 
Jerry, Danny, Elliot
 
Jerry, Danny, Elliot
 +
<p>
 
3A Assembly, Transformation, Agarose Gel Electrophoresis
 
3A Assembly, Transformation, Agarose Gel Electrophoresis
 +
<p>
 
Conducted a 3A with the following components: BBa_J4450 (pSB1C3), BBa_J61100, BBa_K1033916 #1/2. Ran a 1% digest gel with the following specifications: 1: 1 kb ladder; 2: BBa_K1033916 #1, undigested; 3: BBa_K1033916 #1, EcoRI digested; 4: BBa_K1033916 #1, SpeI digested; 5: BBa_K1033916 #1, double digest; 6: BBa_K1033916 #2, undigested; 7: BBa_K1033916 #2, EcoRI digested; 8: BBa_K1033916 #2, SpeI digested; 9: BBa_K1033916 #2, double digest; 10: BBa_K861171 #1, undigested; 11: BBa_K861171 #1, EcoRI digested; 12: BBa_K861171 #1, SpeI digested; 13: BBa_K861171 #1, double digest; 14: 100 bp ladder.
 
Conducted a 3A with the following components: BBa_J4450 (pSB1C3), BBa_J61100, BBa_K1033916 #1/2. Ran a 1% digest gel with the following specifications: 1: 1 kb ladder; 2: BBa_K1033916 #1, undigested; 3: BBa_K1033916 #1, EcoRI digested; 4: BBa_K1033916 #1, SpeI digested; 5: BBa_K1033916 #1, double digest; 6: BBa_K1033916 #2, undigested; 7: BBa_K1033916 #2, EcoRI digested; 8: BBa_K1033916 #2, SpeI digested; 9: BBa_K1033916 #2, double digest; 10: BBa_K861171 #1, undigested; 11: BBa_K861171 #1, EcoRI digested; 12: BBa_K861171 #1, SpeI digested; 13: BBa_K861171 #1, double digest; 14: 100 bp ladder.
 
</p>
 
</p>
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<p>
 
<p>
 
Connor, Sean
 
Connor, Sean
 +
<p>
 
PCR, DNA Clean + Concentration Prep, Simple Recombination, Liquid Culture
 
PCR, DNA Clean + Concentration Prep, Simple Recombination, Liquid Culture
 +
<p>
 
Redid PCR amplification for MLC parts and prepped the PCR mix with the concentrator kit. Conducted the following simple recombination: pSB1C3 + MLC1/2/3/4. Made a liquid culture for the BBa_K1033916/BBa_J61100/pSB1C3 plasmid.  
 
Redid PCR amplification for MLC parts and prepped the PCR mix with the concentrator kit. Conducted the following simple recombination: pSB1C3 + MLC1/2/3/4. Made a liquid culture for the BBa_K1033916/BBa_J61100/pSB1C3 plasmid.  
 
</p>
 
</p>
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<p>
 
<p>
 
Sean
 
Sean
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Transformed the simple recombination results (MLC1-4 + backbone).
 
Transformed the simple recombination results (MLC1-4 + backbone).
 
</p>
 
</p>
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<p>
 
<p>
 
Sean
 
Sean
 +
<p>
 
Transformation, Liquid Cultures
 
Transformation, Liquid Cultures
 +
<p>
 
Retransformed the simple recombination results due to bad colony growth. Made liquid cultures for MLC1, MLC2 (#1 and #2) and MLC 3.  
 
Retransformed the simple recombination results due to bad colony growth. Made liquid cultures for MLC1, MLC2 (#1 and #2) and MLC 3.  
 +
<p>
 
Elliot
 
Elliot
 +
<p>
 
Agarose Gel Electrophoresis
 
Agarose Gel Electrophoresis
 +
<p>
 
Ran a 1% gel with the following specifications: 1: 1 kb ladder; 2: BBa_E1010#1/BBa_J61100/pSB1C3, undigested; 3: BBa_E1010#1/BBa_J61100/pSB1C3, EcoRI digestion; 4: BBa_E1010#1/BBa_J61100/pSB1C3, SpeI digestion; 5: BBa_E1010#1/BBa_J61100/pSB1C3, double digest; 6: BBa_E1010#3/BBa_J61100/pSB1C3, undigested; 7: BBa_E1010#3/BBa_J61100/pSB1C3, EcoRI digestion; 8: BBa_E1010#3/BBa_J61100/pSB1C3, SpeI digestion; 9: BBa_E1010#3/BBa_J61100/pSB1C3, double digest; 10: 1 kb ladder.
 
Ran a 1% gel with the following specifications: 1: 1 kb ladder; 2: BBa_E1010#1/BBa_J61100/pSB1C3, undigested; 3: BBa_E1010#1/BBa_J61100/pSB1C3, EcoRI digestion; 4: BBa_E1010#1/BBa_J61100/pSB1C3, SpeI digestion; 5: BBa_E1010#1/BBa_J61100/pSB1C3, double digest; 6: BBa_E1010#3/BBa_J61100/pSB1C3, undigested; 7: BBa_E1010#3/BBa_J61100/pSB1C3, EcoRI digestion; 8: BBa_E1010#3/BBa_J61100/pSB1C3, SpeI digestion; 9: BBa_E1010#3/BBa_J61100/pSB1C3, double digest; 10: 1 kb ladder.
 
</p>
 
</p>
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<p>
 
<p>
 
Jerry  
 
Jerry  
 +
<p>
 
Agarose Gel Electrophoresis
 
Agarose Gel Electrophoresis
 +
<p>
 
Ran a 1% gel with the following specifications: 1: 100 bp ladder; 2: MLC1; 3: MLC2; 4: MLC3; 5: MLC4; 6: MLC2#1/BBa_J61100/pSB1C3; 7: MLC2#2/BBa_J61100/pSB1C3; 8: MLC3/BBa_J61100/pSB1C3; 9: 1 kb ladder.
 
Ran a 1% gel with the following specifications: 1: 100 bp ladder; 2: MLC1; 3: MLC2; 4: MLC3; 5: MLC4; 6: MLC2#1/BBa_J61100/pSB1C3; 7: MLC2#2/BBa_J61100/pSB1C3; 8: MLC3/BBa_J61100/pSB1C3; 9: 1 kb ladder.
 
</p>
 
</p>
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<p>
 
<p>
 
Jerry
 
Jerry
 +
<p>
 
3A Assembly
 
3A Assembly
 +
<p>
 
Conducted a 3A assembly with the following components: BBa_J4450 (pSB1C3), BBa_K1033916#2+BBa_J61100/BBa_K592009+BBa_J61100, BBa_K861171/ BBa_K118011. Only managed to get to the digestion step, stored for Connors’ usage tomorrow.
 
Conducted a 3A assembly with the following components: BBa_J4450 (pSB1C3), BBa_K1033916#2+BBa_J61100/BBa_K592009+BBa_J61100, BBa_K861171/ BBa_K118011. Only managed to get to the digestion step, stored for Connors’ usage tomorrow.
 
Connor
 
Connor
 +
<p>
 
Zyppy Plasmid Miniprep
 
Zyppy Plasmid Miniprep
 +
<p>
 
Miniprepped the MLC1/2(1)/2(2)/3+pSB1C3 liquid cultures. Made more LB stock.
 
Miniprepped the MLC1/2(1)/2(2)/3+pSB1C3 liquid cultures. Made more LB stock.
 
</p>
 
</p>
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<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
3A Assembly, Zyppy Plasmid Miniprep
 
3A Assembly, Zyppy Plasmid Miniprep
 +
<p>
 
Finished the ligation step of 3A assembly and stored the plasmids in the freezer. Made glycerol stocks for MLC 1/2/3 (1 is mislabeled as 4!). Miniprepped the MLC1+pSB1C3 construct and nanodropped for concentration determination.
 
Finished the ligation step of 3A assembly and stored the plasmids in the freezer. Made glycerol stocks for MLC 1/2/3 (1 is mislabeled as 4!). Miniprepped the MLC1+pSB1C3 construct and nanodropped for concentration determination.
 
</p>
 
</p>
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<p>
 
<p>
 
Elliot
 
Elliot
 +
<p>
 
ZR Plasmid Miniprep Classic
 
ZR Plasmid Miniprep Classic
 +
<p>
 
Miniprepped 3 BBa_K1033916/BBa_J61100/pSB1C3 construct liquid cultures
 
Miniprepped 3 BBa_K1033916/BBa_J61100/pSB1C3 construct liquid cultures
 
</p>
 
</p>
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<p>
 
<p>
 
Elliot  
 
Elliot  
 +
<p>
 
Liquid Cultures, Transformation
 
Liquid Cultures, Transformation
 +
<p>
 
Made liquid cultures for the BBa_1033916/BBa_J61100/pSB1C3 and the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 constructs. Also plated the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs once more.
 
Made liquid cultures for the BBa_1033916/BBa_J61100/pSB1C3 and the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 constructs. Also plated the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs once more.
 +
<p>
 
Connor
 
Connor
 +
<p>
 
3A Assembly
 
3A Assembly
 +
<p>
 
Constructed a plasmid with the following components: BBa_E1010+BBa_J61100 + BBa_K4450 + MLC1/2(1)/2(2)/3.
 
Constructed a plasmid with the following components: BBa_E1010+BBa_J61100 + BBa_K4450 + MLC1/2(1)/2(2)/3.
 
</p>
 
</p>
Line 253: Line 345:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
Transformation/Liquid Culture
 
Transformation/Liquid Culture
 +
<p>
 
Transformed the 3A assembly results from yesterday (BBa_E1010/BBa_J61100 + BBa_K4450 + MLC1/2(1)/2(2)/3). Also made liquid cultures of the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs.  
 
Transformed the 3A assembly results from yesterday (BBa_E1010/BBa_J61100 + BBa_K4450 + MLC1/2(1)/2(2)/3). Also made liquid cultures of the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs.  
 
*NOTE: plates were overgrown during selection!
 
*NOTE: plates were overgrown during selection!
Line 260: Line 354:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
3A Assembly/Transformation/Liquid Culture
 
3A Assembly/Transformation/Liquid Culture
 +
<p>
 
Attempted another ligation with the backbone (BBa_J4450) and MLC4 digests. Transformed the result. Made liquid cultures of MLC composite parts (BBa_E1010/BBa_J61100 + BBa_K4450 + MLC1/2(1)/2(2)/3).  
 
Attempted another ligation with the backbone (BBa_J4450) and MLC4 digests. Transformed the result. Made liquid cultures of MLC composite parts (BBa_E1010/BBa_J61100 + BBa_K4450 + MLC1/2(1)/2(2)/3).  
 +
<p>
 
*Note: The MLC4 ligation culture failed to grow.
 
*Note: The MLC4 ligation culture failed to grow.
 +
<p>
 
Elliot
 
Elliot
 +
<p>
 
ZR Plasmid Miniprep Classic
 
ZR Plasmid Miniprep Classic
 +
<p>
 
Miniprepped the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs. Nanodropped and found no DNA, so discarded.
 
Miniprepped the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs. Nanodropped and found no DNA, so discarded.
 +
<p>
 
*Note: Liquid cultures had a cellular deposit on the bottom for some reason
 
*Note: Liquid cultures had a cellular deposit on the bottom for some reason
 
</p>
 
</p>
Line 271: Line 372:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
ZR Plasmid Miniprep Classic/Liquid Cultures
 
ZR Plasmid Miniprep Classic/Liquid Cultures
 +
<p>
 
Miniprepped the MLC Composite Parts (BBa_E1010/BBa_J61100 + BBa_K4450 + MLC1/2(1)/2(2)/3). Nanodropped and stored the results. Made liquid cultures of the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs
 
Miniprepped the MLC Composite Parts (BBa_E1010/BBa_J61100 + BBa_K4450 + MLC1/2(1)/2(2)/3). Nanodropped and stored the results. Made liquid cultures of the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs
 
</p>
 
</p>
Line 277: Line 380:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
Simple Recombination/Agarose Gel Electrophoresis/Transformation
 
Simple Recombination/Agarose Gel Electrophoresis/Transformation
 +
<p>
 
Attempted simple recombination with MLC4 and pSB1C3. Transformed the result. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. MLC1/BBa_E1010/pSB1C3 (E+P cut); 4. MLC2(1)/BBa_E1010/pSB1C3 (E+P cut); 5. MLC2(2)/BBa_E1010/pSB1C3 (E+P cut); 6. MLC3/BBa_E1010/pSB1C3 (E+P cut).
 
Attempted simple recombination with MLC4 and pSB1C3. Transformed the result. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. MLC1/BBa_E1010/pSB1C3 (E+P cut); 4. MLC2(1)/BBa_E1010/pSB1C3 (E+P cut); 5. MLC2(2)/BBa_E1010/pSB1C3 (E+P cut); 6. MLC3/BBa_E1010/pSB1C3 (E+P cut).
 
</p>
 
</p>
Line 283: Line 388:
 
<p>
 
<p>
 
Connor and Jerry
 
Connor and Jerry
 +
<p>
 
ZR Plasmid Miniprep Classic/Agarose Gel Electrophoresis
 
ZR Plasmid Miniprep Classic/Agarose Gel Electrophoresis
 +
<p>
 
Miniprepped liquid cultures from the red and white colonie on the MLC Composite Parts plates and the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. MLC1/BBa_E1010/pSB1C3 (E+P cut) (Red); 4. MLC2(1)/BBa_E1010/pSB1C3 (E+P cut) (Red); 5. MLC2(2)/BBa_E1010/pSB1C (E+P cut) (Red)3; 6. BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 (E+P cut); 7. BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 (E+P cut); 8. MLC1/BBa_E1010/pSB1C3 (E+P cut) (White); 4. MLC2(1)/BBa_E1010/pSB1C3 (E+P cut) (White); 5. MLC2(2)/BBa_E1010/pSB1C (E+P cut) (White).
 
Miniprepped liquid cultures from the red and white colonie on the MLC Composite Parts plates and the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. MLC1/BBa_E1010/pSB1C3 (E+P cut) (Red); 4. MLC2(1)/BBa_E1010/pSB1C3 (E+P cut) (Red); 5. MLC2(2)/BBa_E1010/pSB1C (E+P cut) (Red)3; 6. BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 (E+P cut); 7. BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 (E+P cut); 8. MLC1/BBa_E1010/pSB1C3 (E+P cut) (White); 4. MLC2(1)/BBa_E1010/pSB1C3 (E+P cut) (White); 5. MLC2(2)/BBa_E1010/pSB1C (E+P cut) (White).
 
</p>
 
</p>
Line 289: Line 396:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
Liquid Cultures
 
Liquid Cultures
 +
<p>
 
Made liquid cultures of the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs.  
 
Made liquid cultures of the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs.  
 
</p>
 
</p>
Line 295: Line 404:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
ZR Plasmid Miniprep Classic
 
ZR Plasmid Miniprep Classic
 +
<p>
 
Miniprepped the BBa_K118011, BBa_K861171, BBa_K592009, BBa_K1033916, BBa_E1010 and BBa_E1010/BBa_J61100 constructs and nanodropped. Made glycerol stocks of all.
 
Miniprepped the BBa_K118011, BBa_K861171, BBa_K592009, BBa_K1033916, BBa_E1010 and BBa_E1010/BBa_J61100 constructs and nanodropped. Made glycerol stocks of all.
 
</p>
 
</p>
Line 301: Line 412:
 
<p>
 
<p>
 
Connor and Jerry
 
Connor and Jerry
 +
<p>
 
Liquid Culture/Transformation
 
Liquid Culture/Transformation
 +
<p>
 
Transformed BBa_B0034 from plate 4, well 1N with 10 uL USP water. Transformed and plated with AmpR plates.  Made 4 liquid cultures with 1g/L glucose concentration and the MLC Composite Parts to check for chromoprotein expression via glucose induction.  
 
Transformed BBa_B0034 from plate 4, well 1N with 10 uL USP water. Transformed and plated with AmpR plates.  Made 4 liquid cultures with 1g/L glucose concentration and the MLC Composite Parts to check for chromoprotein expression via glucose induction.  
 
</p>
 
</p>
Line 307: Line 420:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
Liquid Culture/Simple Recombination/Transformation
 
Liquid Culture/Simple Recombination/Transformation
 +
<p>
 
Made liquid cultures of BBa_B0034 and BBa_J61100. Attempted simple recombination with MLC4(1)/MLC4(2) and pSB1C3. Transformed the ligation result.  
 
Made liquid cultures of BBa_B0034 and BBa_J61100. Attempted simple recombination with MLC4(1)/MLC4(2) and pSB1C3. Transformed the ligation result.  
 
</p>
 
</p>
Line 313: Line 428:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
ZR Plasmid Miniprep Classic/Liquid Cultures
 
ZR Plasmid Miniprep Classic/Liquid Cultures
 +
<p>
 
Miniprepped the BBa_J61100 and BBa_B0034. Nanodropped and made a glycerol stock of both. Made liquid cultures of the MLC4(1)/pSB1C3 and MLC4(2)/pSB1C3 constructs.
 
Miniprepped the BBa_J61100 and BBa_B0034. Nanodropped and made a glycerol stock of both. Made liquid cultures of the MLC4(1)/pSB1C3 and MLC4(2)/pSB1C3 constructs.
 
</p>
 
</p>
Line 319: Line 436:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
ZR Plasmid Miniprep Classic/3A Assembly/Agarose Gel Electrophoresis/Transformation
 
ZR Plasmid Miniprep Classic/3A Assembly/Agarose Gel Electrophoresis/Transformation
 +
<p>
 
Miniprepped the  MLC4(1)/pSB1C3 and MLC4(2)/pSB1C3 constructs. Conducted 3A assembly to make the following parts: BBa_K1033916/BBa_B0034; BBa_K1033916/BBa_J61100; BBa_K592009/BBa_B0034; BBa_K592009/BBa_J61100; BBa_E1010/BBa_B0034; BBa_E1010/BBa_J61100. Transformed the ligation results. Ran a 1% gel with the following specifications: 1. 100 bp ladder; 2. 1 kb ladder; 3. MLC4(1)/pSB1C3; 4. MLC4(2)/pSB1C3; 5. pSB1C3(1); 6.pSB1C3(2).
 
Miniprepped the  MLC4(1)/pSB1C3 and MLC4(2)/pSB1C3 constructs. Conducted 3A assembly to make the following parts: BBa_K1033916/BBa_B0034; BBa_K1033916/BBa_J61100; BBa_K592009/BBa_B0034; BBa_K592009/BBa_J61100; BBa_E1010/BBa_B0034; BBa_E1010/BBa_J61100. Transformed the ligation results. Ran a 1% gel with the following specifications: 1. 100 bp ladder; 2. 1 kb ladder; 3. MLC4(1)/pSB1C3; 4. MLC4(2)/pSB1C3; 5. pSB1C3(1); 6.pSB1C3(2).
 
</p>
 
</p>
Line 325: Line 444:
 
<p>
 
<p>
 
Jerry
 
Jerry
 +
<p>
 
PCR
 
PCR
 +
<p>
 
Amplified the MLC4 fragment
 
Amplified the MLC4 fragment
 
</p>
 
</p>
Line 331: Line 452:
 
<p>
 
<p>
 
Elliot
 
Elliot
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Plated BBa_K1033931, BBa_K1073023, and BBa_K1033929.
 
Plated BBa_K1033931, BBa_K1073023, and BBa_K1033929.
 
</p>
 
</p>
Line 337: Line 460:
 
<p>
 
<p>
 
Connor and Jerry
 
Connor and Jerry
 +
<p>
 
DNA Clean and Concentrator Prep
 
DNA Clean and Concentrator Prep
 +
<p>
 
Prepped the MLC4 PCR results and nanodropped.
 
Prepped the MLC4 PCR results and nanodropped.
 +
<p>
 
Jerry
 
Jerry
 +
<p>
 
Agarose Gel Electrophoresis/PCR
 
Agarose Gel Electrophoresis/PCR
 +
<p>
 
Ran a 1% gel with the following specifications: 1. 100 bp ladder; 2. 1 kb ladder; 3. MLC4; 4. BBa_K1033931; 5. BBa_E1010. Amplified the MLC1-3 fragments with PCR.
 
Ran a 1% gel with the following specifications: 1. 100 bp ladder; 2. 1 kb ladder; 3. MLC4; 4. BBa_K1033931; 5. BBa_E1010. Amplified the MLC1-3 fragments with PCR.
 
</p>
 
</p>
Line 346: Line 474:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
DNA Clean and Concentrator Prep/Agarose Gel Electrophoresis
 
DNA Clean and Concentrator Prep/Agarose Gel Electrophoresis
 +
<p>
 
Prepped the MLC1-3 PCR Results and nanodropped. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. MLC1; 4. MLC2; 5. MLC3; 6. MLC4.
 
Prepped the MLC1-3 PCR Results and nanodropped. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. MLC1; 4. MLC2; 5. MLC3; 6. MLC4.
 +
<p>
 
Jerry/Elliot
 
Jerry/Elliot
 +
<p>
 
Liquid Culture
 
Liquid Culture
 +
<p>
 
Made 2 liquid cultures of BBa_K1073025, BBa_K1073023 and BBa_K1073021 each. Also made liquid cultures of each of the MLC+pSB1C3 constructs.
 
Made 2 liquid cultures of BBa_K1073025, BBa_K1073023 and BBa_K1073021 each. Also made liquid cultures of each of the MLC+pSB1C3 constructs.
 
</p>
 
</p>
Line 355: Line 488:
 
<p>
 
<p>
 
Elliot
 
Elliot
 +
<p>
 
ZR Plasmid Miniprep Classic/3A Assembly
 
ZR Plasmid Miniprep Classic/3A Assembly
 +
<p>
 
Miniprepped BBa_K1033931, BBa_K1033929, and BBa_K1073023. Nanodropped and created the following constructs with 3A: BBa_K118011/BBa_K1073023 and BBa_K861171/BBa_K1033929.  
 
Miniprepped BBa_K1033931, BBa_K1033929, and BBa_K1073023. Nanodropped and created the following constructs with 3A: BBa_K118011/BBa_K1073023 and BBa_K861171/BBa_K1033929.  
 +
<p>
 
Connor
 
Connor
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Transformed the following constructs: BBa_K861171/BBa_K1033929; BBa_K118011/BBa_K1073023; MLC1/pSB1C3; MLC2/pSB1C3; MLC3/pSB1C3; MLC4/pSB1C3.
 
Transformed the following constructs: BBa_K861171/BBa_K1033929; BBa_K118011/BBa_K1073023; MLC1/pSB1C3; MLC2/pSB1C3; MLC3/pSB1C3; MLC4/pSB1C3.
 
</p>
 
</p>
Line 364: Line 502:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
Liquid Culture/Agarose Gel Electrophoresis
 
Liquid Culture/Agarose Gel Electrophoresis
 +
<p>
 
Made liquid cultures for each of the MLC Constructs in backbones. Ran a 1% gel with the following specifications: 1. 100 bp ladder; 2. 1 kb ladder; 3. BBa_K118011/BBa_K1073023 (1) (White); 4. BBa_K118011/BBa_K1073023 (2) (White); 5. BBa_K118011/BBa_K1073023 (1) (Red); 6. BBa_K118011/BBa_K1073023 (2) (Red); 7. BBa_K861171/BBa_K1033929 (1); 8. BBa_K861171/BBa_K1033929 (2).
 
Made liquid cultures for each of the MLC Constructs in backbones. Ran a 1% gel with the following specifications: 1. 100 bp ladder; 2. 1 kb ladder; 3. BBa_K118011/BBa_K1073023 (1) (White); 4. BBa_K118011/BBa_K1073023 (2) (White); 5. BBa_K118011/BBa_K1073023 (1) (Red); 6. BBa_K118011/BBa_K1073023 (2) (Red); 7. BBa_K861171/BBa_K1033929 (1); 8. BBa_K861171/BBa_K1033929 (2).
 
</p>
 
</p>
Line 370: Line 510:
 
<p>
 
<p>
 
Connor/Elliot
 
Connor/Elliot
 +
<p>
 
Agarose Gel Electrophoresis
 
Agarose Gel Electrophoresis
 +
<p>
 
Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. BBa_K118011/BBa_K1073023 (1) (White) (E+P cut); 4. BBa_K118011/BBa_K1073023 (2) (White) (E+P cut); 5. BBa_K118011/BBa_K1073023 (1) (Red) (E+P cut); 6. BBa_K118011/BBa_K1073023 (2) (Red) (E+P cut); 7. BBa_K861171/BBa_K1033929 (1) (E+P cut); 8. BBa_K861171/BBa_K1033929 (2) (E+P cut).
 
Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. BBa_K118011/BBa_K1073023 (1) (White) (E+P cut); 4. BBa_K118011/BBa_K1073023 (2) (White) (E+P cut); 5. BBa_K118011/BBa_K1073023 (1) (Red) (E+P cut); 6. BBa_K118011/BBa_K1073023 (2) (Red) (E+P cut); 7. BBa_K861171/BBa_K1033929 (1) (E+P cut); 8. BBa_K861171/BBa_K1033929 (2) (E+P cut).
 
</p>
 
</p>
Line 376: Line 518:
 
<p>
 
<p>
 
Jerry
 
Jerry
 +
<p>
 
Agarose Gel Electrophoresis
 
Agarose Gel Electrophoresis
 +
<p>
 
Ran a 1% gel with the following specifications (all E+P cut): 1. 1 kb ladder; 2. 100 bp ladder; 3. MLC1/pSB1C3 (a); 4.  MLC1/pSB1C3 (b); 5.  MLC1/pSB1C3 (c); 6.  MLC2/pSB1C3 (b); 7.  MLC2/pSB1C3 (c); 8.  MLC3/pSB1C3 (b); 9.  MLC4/pSB1C3 (a); 10.  MLC4/pSB1C3 (b); 11.  BBa_K861171/BBa_K1033929; 12. BBa_K118011/BBa_K1073023; 13. 100 bp ladder; 14. 1 kb ladder.
 
Ran a 1% gel with the following specifications (all E+P cut): 1. 1 kb ladder; 2. 100 bp ladder; 3. MLC1/pSB1C3 (a); 4.  MLC1/pSB1C3 (b); 5.  MLC1/pSB1C3 (c); 6.  MLC2/pSB1C3 (b); 7.  MLC2/pSB1C3 (c); 8.  MLC3/pSB1C3 (b); 9.  MLC4/pSB1C3 (a); 10.  MLC4/pSB1C3 (b); 11.  BBa_K861171/BBa_K1033929; 12. BBa_K118011/BBa_K1073023; 13. 100 bp ladder; 14. 1 kb ladder.
 
</p>
 
</p>
Line 382: Line 526:
 
<p>
 
<p>
 
Jerry
 
Jerry
 +
<p>
 
Liquid Culture
 
Liquid Culture
 +
<p>
 
Made liquid cultures of BBa_K118011 and BBa_K861171. Also plated the BBa_K1073023,  BBa_K1033929, and BBa_K1033931 sent from the registry.
 
Made liquid cultures of BBa_K118011 and BBa_K861171. Also plated the BBa_K1073023,  BBa_K1033929, and BBa_K1033931 sent from the registry.
−
July 30th, 2015
+
<p>
 +
<h4 style = "font-size: 24 px"> July 30th, 2015</h>
 +
<p>
 
Jerry
 
Jerry
 +
<p>
 +
 
ZR Plasmid Miniprep Classic
 
ZR Plasmid Miniprep Classic
 +
<p>
 +
 
Miniprepped the BBa_K118011 and BBa_K861171 cultures. Nanodropped the result.
 
Miniprepped the BBa_K118011 and BBa_K861171 cultures. Nanodropped the result.
 
</p>
 
</p>
Line 392: Line 544:
 
<p>
 
<p>
 
Jerry
 
Jerry
 +
<p>
 
3A Assembly/Simple Recombination/Agarose Gel Electrophoresis
 
3A Assembly/Simple Recombination/Agarose Gel Electrophoresis
 +
<p>
 +
 
Conducted a 3A to create the following constructs: BBa_K861171/BBa_K1033929 and  BBa_K118011/BBa_K1073023. Attempted to recombine the MLC fragments into the pSB1C3 backbone. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. pSB1C3 (E cut); 4. pSB1C3 (P cut); 5. pSB1C3; 6. MLC1 (E+S cut); 7. MLC2 (E+S cut); 8. MLC3 (E+S cut); 9. MLC4 (E+S cut); 10. pSB1C3 (E+S cut); 11. BBa_K1073023 (P+X cut); 12. BBa_K107029 (P+X cut); 13. BBa_K861171 (P+S cut); 14. BBa_K118011 (P+S cut).
 
Conducted a 3A to create the following constructs: BBa_K861171/BBa_K1033929 and  BBa_K118011/BBa_K1073023. Attempted to recombine the MLC fragments into the pSB1C3 backbone. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. pSB1C3 (E cut); 4. pSB1C3 (P cut); 5. pSB1C3; 6. MLC1 (E+S cut); 7. MLC2 (E+S cut); 8. MLC3 (E+S cut); 9. MLC4 (E+S cut); 10. pSB1C3 (E+S cut); 11. BBa_K1073023 (P+X cut); 12. BBa_K107029 (P+X cut); 13. BBa_K861171 (P+S cut); 14. BBa_K118011 (P+S cut).
 
</p>
 
</p>
Line 398: Line 553:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Transformed all the MLC/pSB1C3 constructs with one of the BBa_K118011/BBa_K1073023 (Red) and BBa_K861171/BBa_K1033929 constructs.
 
Transformed all the MLC/pSB1C3 constructs with one of the BBa_K118011/BBa_K1073023 (Red) and BBa_K861171/BBa_K1033929 constructs.
 
</p>
 
</p>
Line 404: Line 561:
 
<p>
 
<p>
 
Jerry
 
Jerry
 +
<p>
 
Liquid Culture
 
Liquid Culture
 +
<p>
 
Made 2 liquid cultures each of the MLC constructs with pSB1C3,  BBa_K118011/BBa_K1073023, and BBa_K861171/BBa_K1033929.  
 
Made 2 liquid cultures each of the MLC constructs with pSB1C3,  BBa_K118011/BBa_K1073023, and BBa_K861171/BBa_K1033929.  
 
</p>
 
</p>
Line 410: Line 569:
 
<p>
 
<p>
 
Jerry
 
Jerry
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Transformed the reconstituted BBa_J23119 (plate 3, well 17) along with two of the BBa_K118011/BBa_K1073023 constructs and one of the BBa_K861171/BBa_K1033929 constructs.  
 
Transformed the reconstituted BBa_J23119 (plate 3, well 17) along with two of the BBa_K118011/BBa_K1073023 constructs and one of the BBa_K861171/BBa_K1033929 constructs.  
 +
<p>
 
Jerry and Sean
 
Jerry and Sean
 +
<p>
 
ZR Plasmid Miniprep Classic
 
ZR Plasmid Miniprep Classic
 +
<p>
 
Prepped the MLC+pSB1C3, BBa_K118011/BBa_K1073023, and BBa_K861171/BBa_K1033929 constructs. Nanodropped the results.
 
Prepped the MLC+pSB1C3, BBa_K118011/BBa_K1073023, and BBa_K861171/BBa_K1033929 constructs. Nanodropped the results.
 
</p>
 
</p>
Line 419: Line 583:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
3A Assembly/ZR Plasmid Miniprep Classic
 
3A Assembly/ZR Plasmid Miniprep Classic
 +
<p>
 
Attempted a 3A to make the following constructs: MLC1/BBa_K1033931; MLC2/BBa_K1033931; MLC3/BBa_K1033931; MLC4/BBa_K1033931. Miniprepped the pSB1C3 liquid cultures and nanodropped.
 
Attempted a 3A to make the following constructs: MLC1/BBa_K1033931; MLC2/BBa_K1033931; MLC3/BBa_K1033931; MLC4/BBa_K1033931. Miniprepped the pSB1C3 liquid cultures and nanodropped.
 +
<p>
 
Sean
 
Sean
 +
<p>
 
Elongated Digestion/DNA Clean and Concentrator Prep/Elongated Ligation
 
Elongated Digestion/DNA Clean and Concentrator Prep/Elongated Ligation
 +
<p>
 
Digested BBa_J23119, BBa_K1033931, BBa_K1073023, and BBa_K1033929 for 2 hours. Prepped the digestion results and nanodropped. Ligated and left overnight.
 
Digested BBa_J23119, BBa_K1033931, BBa_K1073023, and BBa_K1033929 for 2 hours. Prepped the digestion results and nanodropped. Ligated and left overnight.
 +
<p>
 
Elliot
 
Elliot
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Transformed MLC1-4+pSB1C3.
 
Transformed MLC1-4+pSB1C3.
 
</p>
 
</p>
Line 431: Line 603:
 
<p>
 
<p>
 
Connor/Elliot
 
Connor/Elliot
 +
<p>
 
Liquid Culture/Transformation/Agarose Gel Electrophoresis
 
Liquid Culture/Transformation/Agarose Gel Electrophoresis
 +
<p>
 
Made 4 liquid cultures of the MLC/BBa_K1033931 Constructs. Transformed the constitutive promoter (BBa_J23119)+chromoprotein constructs. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. BBa_K118011/BBa_K1073023 (1); 4. BBa_K118011/BBa_K1073023 (2); 5. BBa_K861171/BBa_K1033929 (1); 6. BBa_K861171/BBa_K1033929 (2).
 
Made 4 liquid cultures of the MLC/BBa_K1033931 Constructs. Transformed the constitutive promoter (BBa_J23119)+chromoprotein constructs. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. BBa_K118011/BBa_K1073023 (1); 4. BBa_K118011/BBa_K1073023 (2); 5. BBa_K861171/BBa_K1033929 (1); 6. BBa_K861171/BBa_K1033929 (2).
 
</p>
 
</p>
Line 437: Line 611:
 
<p>
 
<p>
 
Elliot
 
Elliot
 +
<p>
 
ZR Plasmid Miniprep Classic
 
ZR Plasmid Miniprep Classic
 +
<p>
 
Miniprepped the MLC1-4/pSB1C3/BBa_K1033931 liquid cultures and nanodroppped.
 
Miniprepped the MLC1-4/pSB1C3/BBa_K1033931 liquid cultures and nanodroppped.
 +
<p>
 
Sean
 
Sean
 
DNA Clean and Concentrator Prep/Elongated Ligation
 
DNA Clean and Concentrator Prep/Elongated Ligation
 +
<p>
 
Prepped the digestion results of BBa_K861171, BBa_K118011, BBa_K1033929, and BBa_K1073023. Ligated and left overnight.
 
Prepped the digestion results of BBa_K861171, BBa_K118011, BBa_K1033929, and BBa_K1073023. Ligated and left overnight.
 
</p>
 
</p>
Line 446: Line 624:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Transformed 2 uL of reconstituted pSB1A3 from plate 4, well 2H.
 
Transformed 2 uL of reconstituted pSB1A3 from plate 4, well 2H.
 
</p>
 
</p>
Line 452: Line 632:
 
<p>
 
<p>
 
Connor
 
Connor
 +
<p>
 
Liquid Culture
 
Liquid Culture
 +
<p>
 
Made a liquid culture of pSB1A3.
 
Made a liquid culture of pSB1A3.
 
</p>
 
</p>
Line 458: Line 640:
 
<p>
 
<p>
 
Elliot
 
Elliot
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Transformed 2 uL of reconstituted pSB1A3 from plate 4, well 2H.
 
Transformed 2 uL of reconstituted pSB1A3 from plate 4, well 2H.
 
</p>
 
</p>
Line 464: Line 648:
 
<p>
 
<p>
 
Elliot
 
Elliot
 +
<p>
 
Liquid Culture
 
Liquid Culture
 +
<p>
 
Made a liquid culture of pSB1A3.
 
Made a liquid culture of pSB1A3.
 
</p>
 
</p>
Line 470: Line 656:
 
<p>
 
<p>
 
Jerry/Sean
 
Jerry/Sean
 +
<p>
 
Elongated Digestion/Agarose Gel Electrophoresis
 
Elongated Digestion/Agarose Gel Electrophoresis
 +
<p>
 
Digested MLC+pSB1C3, BBa_K861171, and BBa_K118011 at S+P. Digested BBa_K1033929, BBa_K1073023 and BBa_K1033931 at X+P. Digested for 3 hours in total. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. BBa_K1073023; 3. BBa_K1033929; 4. BBa_K1033931; 5. BBa_K118011; 6. BBa_K861171; 7. MLC1+pSB1C3; 8. MLC2+pSB1C3; 9. MLC+pSB1C3; 10. BBa_J23119.
 
Digested MLC+pSB1C3, BBa_K861171, and BBa_K118011 at S+P. Digested BBa_K1033929, BBa_K1073023 and BBa_K1033931 at X+P. Digested for 3 hours in total. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. BBa_K1073023; 3. BBa_K1033929; 4. BBa_K1033931; 5. BBa_K118011; 6. BBa_K861171; 7. MLC1+pSB1C3; 8. MLC2+pSB1C3; 9. MLC+pSB1C3; 10. BBa_J23119.
 +
<p>
 
Sean
 
Sean
 +
<p>
 
Gel Purification Prep/Elongated Ligation
 
Gel Purification Prep/Elongated Ligation
 +
<p>
 
Ran the digestion results on a gel and purified. Nanodropped and made ligation mixes for 3 hour incubation.
 
Ran the digestion results on a gel and purified. Nanodropped and made ligation mixes for 3 hour incubation.
 +
<p>
 
Elliot
 
Elliot
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Transformed the final ligation mixtures, including BBa_K118011/BBa_K1073023, BBa_K861171/BBa_K1033931, and MLC1-4/BBa_K1033929.
 
Transformed the final ligation mixtures, including BBa_K118011/BBa_K1073023, BBa_K861171/BBa_K1033931, and MLC1-4/BBa_K1033929.
 
</p>
 
</p>
Line 482: Line 676:
 
<p>
 
<p>
 
Elliot
 
Elliot
 +
<p>
 
Transformation
 
Transformation
 +
<p>
 
Retransformed the final ligation mixtures, including BBa_K118011/BBa_K1073023, BBa_K861171/BBa_K1033931, and MLC1-4/BBa_K1033929.
 
Retransformed the final ligation mixtures, including BBa_K118011/BBa_K1073023, BBa_K861171/BBa_K1033931, and MLC1-4/BBa_K1033929.
 
</p>
 
</p>
Line 488: Line 684:
 
<p>
 
<p>
 
Jerry and Sean
 
Jerry and Sean
 +
<p>
 
Elongated Digestion
 
Elongated Digestion
 +
<p>
 
Digested MLC+pSB1C3, BBa_K861171, and BBa_K118011 at S+P. Digested BBa_K1033929, BBa_K1073023 and BBa_K1033931 at X+P. Digested for 3 hours in total.  
 
Digested MLC+pSB1C3, BBa_K861171, and BBa_K118011 at S+P. Digested BBa_K1033929, BBa_K1073023 and BBa_K1033931 at X+P. Digested for 3 hours in total.  
 
</p>
 
</p>
Line 494: Line 692:
 
<p>
 
<p>
 
Sean
 
Sean
 +
<p>
 
DNA Clean and Concentrator/Gel Purification Prep/Elongated Ligation
 
DNA Clean and Concentrator/Gel Purification Prep/Elongated Ligation
 +
<p>
 
Prepped the promoter parts with the concentrator kit and selected for the correct chromoproteins with the gel purification prep. Ligated BBa_J23119 and BBa_K118011 with BBa_K1073023 and left for 3 hours.  
 
Prepped the promoter parts with the concentrator kit and selected for the correct chromoproteins with the gel purification prep. Ligated BBa_J23119 and BBa_K118011 with BBa_K1073023 and left for 3 hours.  
 +
<p>
 
Connor
 
Connor
 +
<p>
 
Transformation/Liquid Culture
 
Transformation/Liquid Culture
 +
<p>
 
Transformed the red chromoprotein+constitutive promoter construct and the BBa_K118011/BBa_K1073023 construct. Made liquid cultures of all of the MLC/pSB1C3 constructs, BBa_1033931, and BBa_K1033929.  
 
Transformed the red chromoprotein+constitutive promoter construct and the BBa_K118011/BBa_K1073023 construct. Made liquid cultures of all of the MLC/pSB1C3 constructs, BBa_1033931, and BBa_K1033929.  
 
</p>
 
</p>

Revision as of 21:51, 17 September 2015

NOTEBOOK

May June July August September

Notebook

May 18th, 2015

Connor and Sean

Transformation

Resuspended parts BBa_E1010 and BBa_I716120 on plate 3 well 11n and plate 3 well 11m, respectively. Transformed both and placed in 37 degree incubator *Note: parts left in 37 degree incubator too long (>24 hours) and overgrew

May 21st, 2015

May 21st, 2015

Elliot and Danny

Transformation

Resuspended part BBa_K118011 on plate 3 well 20c. Transformed this part along with BBa_E1010 again.

June 3rd, 2015

Connor and Sean

Transformation

Resuspended parts BBa_K1033916 and BBa_K592009 on plate 4 well 6m and plate 1 well 15e, respectively. Transformed those parts and BBa_E1010.

*Note: BBa_K1033916 plate was thrown out due to suspicion of LBKanR mislabeling and the two other chromoproteins appear to have been switched.

Elliot and Danny

Transformation

Transformed parts BBa_E1010, BBa_K1033916, BBa_K592009, BBa_K861171 and BBa_K118011.

Will and Jerry

ZR Plasmid Miniprep Classic

Miniprepped parts BBa_K861171 and BBa_K118011. Prepared 1x TAE Buffer from stock.

June 4th, 2015

Will

Transformation

Did overall inventory and plated BBa_K861171

Connor and Sean

Liquid Cultures

Created liquid cultures of BBa_E1010 (2), BBa_K592009 (2), and BBa_K1033916 and placed in 37 degree shaking incubator overnight.

*Note: Liquid cultures spent >24 hours in 37 degree incubator, so thrown out. Also, BBa_K118011 failed to grow.

June 5th, 2015

Elliot and Danny

Liquid Cultures

Created liquid cultures of BBa_E1010 (2), BBa_K592009 (2), and BBa_K1033916 and placed in 37 degree shaking incubator overnight. LB was autoclaved and placed in 4 degree fridge.

June 6th, 2015

Elliot and Danny

ZR Plasmid Miniprep Classic

Miniprepped BBa_E1010 (2), BBa_K592009 (2), and BBa_K1033916 and nanodropped for concentrations.

June 8th, 2015

Will

Did overall inventory and made a 1L DI water stock

June 10th, 2015

Will and Jerry

PCR

Received and reconstituted MLC Parts and MLC primers. Performed PCR amplification on the MLC parts.

June 11th, 2015

Will and Jerry

Agarose Gel Electrophoresis, DNA Clean + Concentrator Prep, PCR

Ran a gel verifying the MLC parts, concentrated the PCR-generated plasmids and nanodropped. After determining that concentrations were insufficient, ran another PCR for MLC amplification. Also conducted inventory.

Connor and Sean

DNA Clean + Concentrator Prep, Transformation

Prepped the DNA and transformed BBa_J4450, which was reconstituted from plate 4 well 4b. Made a glycerol stock.

Elliot and Danny

Transformation

Created ampicillin plates and reconstituted BBa_J61100 on plate 4 well b4. Transformed BBa_J61100 on the ampicillin plates.

June 12th, 2015

Connor, Sean and Elliot

Liquid Cultures, DNA Clean and Concentrator Prep

Made 2 liquid cultures of BBa_J61100 (LBAmp) and 4 liquid cultures of BBa_J4450 (LBCam). Prepped the new MLC PCR mixes and nanodropped for concentrations.

June 13th, 2015

Elliot and Danny

ZR Plasmid Miniprep Classic

Miniprepped the liquid cultures of BBa_J61100 and BBa_J4450 (6 in total). Stored in the -20 degrees freezer.

June 14th, 2015

Elliot and Danny

Liquid Cultures

Created 3 liquid cultures of BBa_J4450 (LBCam) and 6 liquid cultures of BBa_J61100 (LBAmp).

June 15th, 2015

Connor

ZR Plasmid Miniprep Classic

Miniprepped 2 BBa_J4450 cultures and 5 BBa_J61100 cultures. Nanodropped and stored in the -20 degrees freezer.

Elliot and Danny

3A Assembly, Agarose Gel Electrophoresis

Conducted 3A Assembly with the following components: pSB1C3 (from BBa_J4450), BBa_K1033916/BBa_E1010/BBa_K592009 (downstream component) and BBa_J61100 (upstream component). Made a 1% agarose gel with the following wells: 1: 1 kb ladder; 2: BBa_E1010 #3; 3. BBa_E1010 #2; 4. BBa_K1033916 #3; 5. BBa_K592009 #2; 6. BBa_K861171 # 3; 7. 1 kb ladder. Made another 1% agarose gel: 1: 1 kb ladder; 2: 100 bp ladder; 3. BBa_K1033916 #3; 4. BBa_K592009 #2; 5. BBa_K1033916 #1; 6. BBa_K861171 #1; 7. BBa_K861171 #2; 8. BBa_K861171 #3

June 16th, 2015

Elliot + Danny + Jerry

Agarose Gel Electrophoresis

Ran a diagnostic 2% gel for the repressible.

Will + Jerry

Agarose Gel Electrophoresis

Ran a 1% gel with the following specifications: 1: 1 kb ladder; 2: 100 bp ladder; 3: BBa_K1033916 #1; 4: BBa_K1033916 #2; 5: BBa_K1033916 #3; 6: BBa_K592009 #2; 7: BBa_K861171 #1; 8: BBa_K861171 #2; 9: BBa_K861171 #3

Connor + Will

3A Assembly, Simple Recombination

Conducted a 3A assembly with the following components: pSB1C3, BBa_E1010/BBa_K592009, BBa_J1100. Conducted simple recombination (cutting a backbone and “part” with the same enzymes) with the following components: MLC1/2/3/4, pSB1C3.

Will

Transformation

Transformed the two 3A plasmids and four simple recombination plasmids.

*Note: all MLC plates failed to grow

June 17th, 2015

Will

PCR, Transformation

Conducted another PCR amplification procedure for the order MLC parts (1-4). Transformed the MLC + pSB1C3 constructs that failed to grow once more.

June 18th, 2015

Will

Agarose Gel Electrophoresis, Gel DNA Recovery Prep

Made glycerol stocks of BBa_K1033916#3/pSB1C3/BBa_J61100 (pink, so actually BBa_J4450), BBa_K592009#2/pSB1C3/BBa_J61100, BBa_E1010#2/pSB1C3/BBa_J61100. Then ran a 1% gel with the following specifications: 1: 1 kb ladder; 2: 100 bp ladder; 3: MLC1: 4: MLC2; 5: MLC3; 6: MLC4; 7: MasterMix; 8: USP H20. Then ran a DNA gel recovery prep and nanodropped. *Note: MLC remainders from the gel were not enough, so another PCR is recommended.

Connor

Zyppy Plasmid Miniprep, Transformation

Miniprepped BBa_K592009/BBa_J61100/pSB1C3 and BBa_E1010/BBa_J61100/pSB1C3 liquid cultures. Nanodropped the results. Transformed the BBa_1033916 #1/#2 3A results.

Jerry, Danny, Elliot

3A Assembly, Transformation, Agarose Gel Electrophoresis

Conducted a 3A with the following components: BBa_J4450 (pSB1C3), BBa_J61100, BBa_K1033916 #1/2. Ran a 1% digest gel with the following specifications: 1: 1 kb ladder; 2: BBa_K1033916 #1, undigested; 3: BBa_K1033916 #1, EcoRI digested; 4: BBa_K1033916 #1, SpeI digested; 5: BBa_K1033916 #1, double digest; 6: BBa_K1033916 #2, undigested; 7: BBa_K1033916 #2, EcoRI digested; 8: BBa_K1033916 #2, SpeI digested; 9: BBa_K1033916 #2, double digest; 10: BBa_K861171 #1, undigested; 11: BBa_K861171 #1, EcoRI digested; 12: BBa_K861171 #1, SpeI digested; 13: BBa_K861171 #1, double digest; 14: 100 bp ladder.

June 19th, 2015

Connor, Sean

PCR, DNA Clean + Concentration Prep, Simple Recombination, Liquid Culture

Redid PCR amplification for MLC parts and prepped the PCR mix with the concentrator kit. Conducted the following simple recombination: pSB1C3 + MLC1/2/3/4. Made a liquid culture for the BBa_K1033916/BBa_J61100/pSB1C3 plasmid.

June 20th, 2015

Sean

Transformation

Transformed the simple recombination results (MLC1-4 + backbone).

June 21th, 2015

Sean

Transformation, Liquid Cultures

Retransformed the simple recombination results due to bad colony growth. Made liquid cultures for MLC1, MLC2 (#1 and #2) and MLC 3.

Elliot

Agarose Gel Electrophoresis

Ran a 1% gel with the following specifications: 1: 1 kb ladder; 2: BBa_E1010#1/BBa_J61100/pSB1C3, undigested; 3: BBa_E1010#1/BBa_J61100/pSB1C3, EcoRI digestion; 4: BBa_E1010#1/BBa_J61100/pSB1C3, SpeI digestion; 5: BBa_E1010#1/BBa_J61100/pSB1C3, double digest; 6: BBa_E1010#3/BBa_J61100/pSB1C3, undigested; 7: BBa_E1010#3/BBa_J61100/pSB1C3, EcoRI digestion; 8: BBa_E1010#3/BBa_J61100/pSB1C3, SpeI digestion; 9: BBa_E1010#3/BBa_J61100/pSB1C3, double digest; 10: 1 kb ladder.

June 22nd, 2015

Jerry

Agarose Gel Electrophoresis

Ran a 1% gel with the following specifications: 1: 100 bp ladder; 2: MLC1; 3: MLC2; 4: MLC3; 5: MLC4; 6: MLC2#1/BBa_J61100/pSB1C3; 7: MLC2#2/BBa_J61100/pSB1C3; 8: MLC3/BBa_J61100/pSB1C3; 9: 1 kb ladder.

June 23rd, 2015

Jerry

3A Assembly

Conducted a 3A assembly with the following components: BBa_J4450 (pSB1C3), BBa_K1033916#2+BBa_J61100/BBa_K592009+BBa_J61100, BBa_K861171/ BBa_K118011. Only managed to get to the digestion step, stored for Connors’ usage tomorrow. Connor

Zyppy Plasmid Miniprep

Miniprepped the MLC1/2(1)/2(2)/3+pSB1C3 liquid cultures. Made more LB stock.

June 24th, 2015

Connor

3A Assembly, Zyppy Plasmid Miniprep

Finished the ligation step of 3A assembly and stored the plasmids in the freezer. Made glycerol stocks for MLC 1/2/3 (1 is mislabeled as 4!). Miniprepped the MLC1+pSB1C3 construct and nanodropped for concentration determination.

June 26th, 2015

Elliot

ZR Plasmid Miniprep Classic

Miniprepped 3 BBa_K1033916/BBa_J61100/pSB1C3 construct liquid cultures

July 1st, 2015

Elliot

Liquid Cultures, Transformation

Made liquid cultures for the BBa_1033916/BBa_J61100/pSB1C3 and the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 constructs. Also plated the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs once more.

Connor

3A Assembly

Constructed a plasmid with the following components: BBa_E1010+BBa_J61100 + BBa_K4450 + MLC1/2(1)/2(2)/3.

July 2nd, 2015

Connor

Transformation/Liquid Culture

Transformed the 3A assembly results from yesterday (BBa_E1010/BBa_J61100 + BBa_K4450 + MLC1/2(1)/2(2)/3). Also made liquid cultures of the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs. *NOTE: plates were overgrown during selection!

July 3rd, 2015

Connor

3A Assembly/Transformation/Liquid Culture

Attempted another ligation with the backbone (BBa_J4450) and MLC4 digests. Transformed the result. Made liquid cultures of MLC composite parts (BBa_E1010/BBa_J61100 + BBa_K4450 + MLC1/2(1)/2(2)/3).

*Note: The MLC4 ligation culture failed to grow.

Elliot

ZR Plasmid Miniprep Classic

Miniprepped the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs. Nanodropped and found no DNA, so discarded.

*Note: Liquid cultures had a cellular deposit on the bottom for some reason

July 4th, 2015

Connor

ZR Plasmid Miniprep Classic/Liquid Cultures

Miniprepped the MLC Composite Parts (BBa_E1010/BBa_J61100 + BBa_K4450 + MLC1/2(1)/2(2)/3). Nanodropped and stored the results. Made liquid cultures of the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs

July 6th, 2015

Connor

Simple Recombination/Agarose Gel Electrophoresis/Transformation

Attempted simple recombination with MLC4 and pSB1C3. Transformed the result. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. MLC1/BBa_E1010/pSB1C3 (E+P cut); 4. MLC2(1)/BBa_E1010/pSB1C3 (E+P cut); 5. MLC2(2)/BBa_E1010/pSB1C3 (E+P cut); 6. MLC3/BBa_E1010/pSB1C3 (E+P cut).

July 7th, 2015

Connor and Jerry

ZR Plasmid Miniprep Classic/Agarose Gel Electrophoresis

Miniprepped liquid cultures from the red and white colonie on the MLC Composite Parts plates and the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. MLC1/BBa_E1010/pSB1C3 (E+P cut) (Red); 4. MLC2(1)/BBa_E1010/pSB1C3 (E+P cut) (Red); 5. MLC2(2)/BBa_E1010/pSB1C (E+P cut) (Red)3; 6. BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 (E+P cut); 7. BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 (E+P cut); 8. MLC1/BBa_E1010/pSB1C3 (E+P cut) (White); 4. MLC2(1)/BBa_E1010/pSB1C3 (E+P cut) (White); 5. MLC2(2)/BBa_E1010/pSB1C (E+P cut) (White).

July 8th, 2015

Connor

Liquid Cultures

Made liquid cultures of the BBa_K118011/BBa_K592009+BBa_J61100/pSB1C3 and BBa_K861171/BBa_1033916+BBa_J61100/pSB1C3 constructs.

July 9th, 2015

Connor

ZR Plasmid Miniprep Classic

Miniprepped the BBa_K118011, BBa_K861171, BBa_K592009, BBa_K1033916, BBa_E1010 and BBa_E1010/BBa_J61100 constructs and nanodropped. Made glycerol stocks of all.

July 10th, 2015

Connor and Jerry

Liquid Culture/Transformation

Transformed BBa_B0034 from plate 4, well 1N with 10 uL USP water. Transformed and plated with AmpR plates. Made 4 liquid cultures with 1g/L glucose concentration and the MLC Composite Parts to check for chromoprotein expression via glucose induction.

July 11th, 2015

Connor

Liquid Culture/Simple Recombination/Transformation

Made liquid cultures of BBa_B0034 and BBa_J61100. Attempted simple recombination with MLC4(1)/MLC4(2) and pSB1C3. Transformed the ligation result.

July 12th, 2015

Connor

ZR Plasmid Miniprep Classic/Liquid Cultures

Miniprepped the BBa_J61100 and BBa_B0034. Nanodropped and made a glycerol stock of both. Made liquid cultures of the MLC4(1)/pSB1C3 and MLC4(2)/pSB1C3 constructs.

July 13th, 2015

Connor

ZR Plasmid Miniprep Classic/3A Assembly/Agarose Gel Electrophoresis/Transformation

Miniprepped the MLC4(1)/pSB1C3 and MLC4(2)/pSB1C3 constructs. Conducted 3A assembly to make the following parts: BBa_K1033916/BBa_B0034; BBa_K1033916/BBa_J61100; BBa_K592009/BBa_B0034; BBa_K592009/BBa_J61100; BBa_E1010/BBa_B0034; BBa_E1010/BBa_J61100. Transformed the ligation results. Ran a 1% gel with the following specifications: 1. 100 bp ladder; 2. 1 kb ladder; 3. MLC4(1)/pSB1C3; 4. MLC4(2)/pSB1C3; 5. pSB1C3(1); 6.pSB1C3(2).

July 14th, 2015

Jerry

PCR

Amplified the MLC4 fragment

July 15th, 2015

Elliot

Transformation

Plated BBa_K1033931, BBa_K1073023, and BBa_K1033929.

July 16th, 2015

Connor and Jerry

DNA Clean and Concentrator Prep

Prepped the MLC4 PCR results and nanodropped.

Jerry

Agarose Gel Electrophoresis/PCR

Ran a 1% gel with the following specifications: 1. 100 bp ladder; 2. 1 kb ladder; 3. MLC4; 4. BBa_K1033931; 5. BBa_E1010. Amplified the MLC1-3 fragments with PCR.

July 17th, 2015

Connor

DNA Clean and Concentrator Prep/Agarose Gel Electrophoresis

Prepped the MLC1-3 PCR Results and nanodropped. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. MLC1; 4. MLC2; 5. MLC3; 6. MLC4.

Jerry/Elliot

Liquid Culture

Made 2 liquid cultures of BBa_K1073025, BBa_K1073023 and BBa_K1073021 each. Also made liquid cultures of each of the MLC+pSB1C3 constructs.

July 18th, 2015

Elliot

ZR Plasmid Miniprep Classic/3A Assembly

Miniprepped BBa_K1033931, BBa_K1033929, and BBa_K1073023. Nanodropped and created the following constructs with 3A: BBa_K118011/BBa_K1073023 and BBa_K861171/BBa_K1033929.

Connor

Transformation

Transformed the following constructs: BBa_K861171/BBa_K1033929; BBa_K118011/BBa_K1073023; MLC1/pSB1C3; MLC2/pSB1C3; MLC3/pSB1C3; MLC4/pSB1C3.

July 20th, 2015

Connor

Liquid Culture/Agarose Gel Electrophoresis

Made liquid cultures for each of the MLC Constructs in backbones. Ran a 1% gel with the following specifications: 1. 100 bp ladder; 2. 1 kb ladder; 3. BBa_K118011/BBa_K1073023 (1) (White); 4. BBa_K118011/BBa_K1073023 (2) (White); 5. BBa_K118011/BBa_K1073023 (1) (Red); 6. BBa_K118011/BBa_K1073023 (2) (Red); 7. BBa_K861171/BBa_K1033929 (1); 8. BBa_K861171/BBa_K1033929 (2).

July 21st, 2015

Connor/Elliot

Agarose Gel Electrophoresis

Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. BBa_K118011/BBa_K1073023 (1) (White) (E+P cut); 4. BBa_K118011/BBa_K1073023 (2) (White) (E+P cut); 5. BBa_K118011/BBa_K1073023 (1) (Red) (E+P cut); 6. BBa_K118011/BBa_K1073023 (2) (Red) (E+P cut); 7. BBa_K861171/BBa_K1033929 (1) (E+P cut); 8. BBa_K861171/BBa_K1033929 (2) (E+P cut).

July 23rd, 2015

Jerry

Agarose Gel Electrophoresis

Ran a 1% gel with the following specifications (all E+P cut): 1. 1 kb ladder; 2. 100 bp ladder; 3. MLC1/pSB1C3 (a); 4. MLC1/pSB1C3 (b); 5. MLC1/pSB1C3 (c); 6. MLC2/pSB1C3 (b); 7. MLC2/pSB1C3 (c); 8. MLC3/pSB1C3 (b); 9. MLC4/pSB1C3 (a); 10. MLC4/pSB1C3 (b); 11. BBa_K861171/BBa_K1033929; 12. BBa_K118011/BBa_K1073023; 13. 100 bp ladder; 14. 1 kb ladder.

July 29th, 2015

Jerry

Liquid Culture

Made liquid cultures of BBa_K118011 and BBa_K861171. Also plated the BBa_K1073023, BBa_K1033929, and BBa_K1033931 sent from the registry.

July 30th, 2015

Jerry

ZR Plasmid Miniprep Classic

Miniprepped the BBa_K118011 and BBa_K861171 cultures. Nanodropped the result.

July 31st, 2015

Jerry

3A Assembly/Simple Recombination/Agarose Gel Electrophoresis

Conducted a 3A to create the following constructs: BBa_K861171/BBa_K1033929 and BBa_K118011/BBa_K1073023. Attempted to recombine the MLC fragments into the pSB1C3 backbone. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. pSB1C3 (E cut); 4. pSB1C3 (P cut); 5. pSB1C3; 6. MLC1 (E+S cut); 7. MLC2 (E+S cut); 8. MLC3 (E+S cut); 9. MLC4 (E+S cut); 10. pSB1C3 (E+S cut); 11. BBa_K1073023 (P+X cut); 12. BBa_K107029 (P+X cut); 13. BBa_K861171 (P+S cut); 14. BBa_K118011 (P+S cut).

August 1st, 2015

Connor

Transformation

Transformed all the MLC/pSB1C3 constructs with one of the BBa_K118011/BBa_K1073023 (Red) and BBa_K861171/BBa_K1033929 constructs.

August 2nd, 2015

Jerry

Liquid Culture

Made 2 liquid cultures each of the MLC constructs with pSB1C3, BBa_K118011/BBa_K1073023, and BBa_K861171/BBa_K1033929.

August 3rd, 2015

Jerry

Transformation

Transformed the reconstituted BBa_J23119 (plate 3, well 17) along with two of the BBa_K118011/BBa_K1073023 constructs and one of the BBa_K861171/BBa_K1033929 constructs.

Jerry and Sean

ZR Plasmid Miniprep Classic

Prepped the MLC+pSB1C3, BBa_K118011/BBa_K1073023, and BBa_K861171/BBa_K1033929 constructs. Nanodropped the results.

August 4th, 2015

Connor

3A Assembly/ZR Plasmid Miniprep Classic

Attempted a 3A to make the following constructs: MLC1/BBa_K1033931; MLC2/BBa_K1033931; MLC3/BBa_K1033931; MLC4/BBa_K1033931. Miniprepped the pSB1C3 liquid cultures and nanodropped.

Sean

Elongated Digestion/DNA Clean and Concentrator Prep/Elongated Ligation

Digested BBa_J23119, BBa_K1033931, BBa_K1073023, and BBa_K1033929 for 2 hours. Prepped the digestion results and nanodropped. Ligated and left overnight.

Elliot

Transformation

Transformed MLC1-4+pSB1C3.

August 5th, 2015

Connor/Elliot

Liquid Culture/Transformation/Agarose Gel Electrophoresis

Made 4 liquid cultures of the MLC/BBa_K1033931 Constructs. Transformed the constitutive promoter (BBa_J23119)+chromoprotein constructs. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. 100 bp ladder; 3. BBa_K118011/BBa_K1073023 (1); 4. BBa_K118011/BBa_K1073023 (2); 5. BBa_K861171/BBa_K1033929 (1); 6. BBa_K861171/BBa_K1033929 (2).

August 6th, 2015

Elliot

ZR Plasmid Miniprep Classic

Miniprepped the MLC1-4/pSB1C3/BBa_K1033931 liquid cultures and nanodroppped.

Sean DNA Clean and Concentrator Prep/Elongated Ligation

Prepped the digestion results of BBa_K861171, BBa_K118011, BBa_K1033929, and BBa_K1073023. Ligated and left overnight.

August 7th, 2015

Connor

Transformation

Transformed 2 uL of reconstituted pSB1A3 from plate 4, well 2H.

August 1th, 2015

Connor

Liquid Culture

Made a liquid culture of pSB1A3.

August 10th, 2015

Elliot

Transformation

Transformed 2 uL of reconstituted pSB1A3 from plate 4, well 2H.

August 11th, 2015

Elliot

Liquid Culture

Made a liquid culture of pSB1A3.

August 12th, 2015

Jerry/Sean

Elongated Digestion/Agarose Gel Electrophoresis

Digested MLC+pSB1C3, BBa_K861171, and BBa_K118011 at S+P. Digested BBa_K1033929, BBa_K1073023 and BBa_K1033931 at X+P. Digested for 3 hours in total. Ran a 1% gel with the following specifications: 1. 1 kb ladder; 2. BBa_K1073023; 3. BBa_K1033929; 4. BBa_K1033931; 5. BBa_K118011; 6. BBa_K861171; 7. MLC1+pSB1C3; 8. MLC2+pSB1C3; 9. MLC+pSB1C3; 10. BBa_J23119.

Sean

Gel Purification Prep/Elongated Ligation

Ran the digestion results on a gel and purified. Nanodropped and made ligation mixes for 3 hour incubation.

Elliot

Transformation

Transformed the final ligation mixtures, including BBa_K118011/BBa_K1073023, BBa_K861171/BBa_K1033931, and MLC1-4/BBa_K1033929.

August 13th, 2015

Elliot

Transformation

Retransformed the final ligation mixtures, including BBa_K118011/BBa_K1073023, BBa_K861171/BBa_K1033931, and MLC1-4/BBa_K1033929.

August 14th, 2015

Jerry and Sean

Elongated Digestion

Digested MLC+pSB1C3, BBa_K861171, and BBa_K118011 at S+P. Digested BBa_K1033929, BBa_K1073023 and BBa_K1033931 at X+P. Digested for 3 hours in total.

August 18th, 2015

Sean

DNA Clean and Concentrator/Gel Purification Prep/Elongated Ligation

Prepped the promoter parts with the concentrator kit and selected for the correct chromoproteins with the gel purification prep. Ligated BBa_J23119 and BBa_K118011 with BBa_K1073023 and left for 3 hours.

Connor

Transformation/Liquid Culture

Transformed the red chromoprotein+constitutive promoter construct and the BBa_K118011/BBa_K1073023 construct. Made liquid cultures of all of the MLC/pSB1C3 constructs, BBa_1033931, and BBa_K1033929.

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