Notebook
Format
Month Day, Year
Name(s) of people who completed work
Protocol(s) used
More detailed description
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 9th, 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.
August 19th, 2015
Connor
ZR Plasmid Miniprep Classic/Liquid Culture
Miniprepped BBa_J23119 and BBa_K1073023. Made liquid cultures of BBa_K118011, BBa_K861171, BBa_K1073023, BBa_K1033931, BBa_K1033929, the MLC1-4+pSB1C3 constructs and BBa_J23119.
August 20th, 2015
Connor
ZR Plasmid Miniprep Classic
Miniprepped BBa_K118011, BBa_K861171, BBa_K1073023, BBa_K1033931, BBa_K1033929, the MLC1-4+pSB1C3 constructs and BBa_J23119 and nanodropped for concentration.
August 21st, 2015
Connor
Elongated Digestion/Gel Purification Prep/Elongated Ligation
Digested 2500 ng of BBa_K118011, BBa_K861171, BBa_K1073023, BBa_K1033931, BBa_K1033929, the MLC1-4+pSB1C3 constructs and BBa_J23119 each for 3 hours. Ran on a 1% gel and purified with the kit. Ligated overnight.
August 22nd, 2015
Connor
Transformation
Transformed the ligation mixes of BBa_K118011, BBa_K861171, BBa_K1073023, BBa_K1033931, BBa_K1033929, the MLC1-4+pSB1C3 constructs and BBa_J23119. Incubated overnight.
August 26th, 2015
Connor
ZR Plasmid Miniprep
Miniprepped BBa_K118011/BBa_K1073023, BBa_K861171/BBa_K1073023, and the MLC2 construct/BBa_K1033931.
Jerry
PCR
Amplified the PCR Constructs with a 32 cycle PCR.
August 27th, 2015
Sean
PCR
Amplified the PCR Constructs with a 35 cycle PCR and left overnight in the freezer.
August 28th, 2015
Sean and Jerry
DNA Clean and Concentrate Prep/Elongated Digestion
Cleaned the completed PCR solutions with the prep. Nanodropped for concentrations. Digested the PCR constructs with pSB1C3 and incubated for three hours.
Connor
DNA Clean and Concentrator Prep/Ligation
Prepped the digested PCR constructs and ligated into the digested pSB1C3 backbone. Incubated overnight.
August 29th, 2015
Connor
Transformation
Transformed the ligated PCR+pSB1C3 constructs. Incubated overnight.
August 30th, 2015
Elliot
Transformation
Transformed pSB1C3 stocks and plated. Incubated overnight.
August 31st, 2015
Connor
PCR/DNA Clean and Concentrator Kit/Elongated Digestion/Elongated Ligation
Conducted a 32 cycle PCR amplification of the PCR constructs. Cleaned with the kit and nanodropped. The results were then digested with miniprepped pSB1C3 and cleaned again after a 2 hour incubation. The PCR constructs were then ligated with the digested pSB1C3 and incubated overnight.
September 1st, 2015
Jerry
Transformation/PCR
Transformed the PCR Construct ligation mixes and incubated overnight. Set a 32 cycle PCR to amplify more of the base PCR constructs.
September 2nd, 2015
Connor
Liquid Culture
Made liquid cultures of the MLC/BBa_K1033931 PCR constructs, the BBa_J23119/BBa_K1073023 PCR construct, the BBa_K861171/BBa_K1073023 PCR construct, the BBa_K118011/BBa_K1033929 PCR construct and the tripart constructs.
September 3rd, 2015
Connor
ZR Plasmid Miniprep Classic
Miniprepped the MLC/BBa_K1033931 PCR constructs, the BBa_J23119/BBa_K1073023 PCR construct, the BBa_K861171/BBa_K1073023 PCR construct, the BBa_K118011/BBa_K1033929 PCR construct and the tripart constructs.
September 4th, 2015
Jerry
ZR Plasmid Miniprep Classic
Miniprepped the viable PCR Construct liquid cultures and nanodropped.
Connor
PCR/DNA Clean and Concentrator Kit/Elongated DIgestion/Elongated Ligation
Conducted a 32 cycle PCR amplification of the PCR constructs. Cleaned with the kit and nanodropped. The results were then digested with miniprepped pSB1C3 and cleaned again after a 2 hour incubation. The PCR constructs were then ligated with the digested pSB1C3 and incubated overnight.
September 5th, 2015
Will
DNA Clean and Concentrator Prep/Elongated Ligation
Prepped the BBa_J23119/BBa_K1033931 and BBa_J23119/BBa_K1033929 PCR constructs. Ligated with digested pSB1C3 and incubated overnight.
Connor
Transformation
Transformed the ligated PCR constructs+pSB1C3. Incubated overnight.
September 6th, 2015
Elliot
Liquid Cultures/Transformation
Created liquid cultures of the viable PCR constructs and transformed the BBa_J23119/BBa_K1033931 and BBa_J23119/BBa_K1033929 PCR constructs. Incubated overnight.
September 7th, 2015
Elliot
ZR Plasmid Miniprep Classic
Miniprepped the liquid cultures of the viable PCR constructs.
September 8th, 2015
Connor
PCR/Transformation
Transformed the the ligated PCR constructs+pSB1C3. Incubated overnight. Also ran a 32 cycle PCR amplification of the BBa_J23119/BBa_K1033931 and BBa_J23119/BBa_K1033929 PCR constructs.
September 10th, 2015
Connor
ZR Plasmid Miniprep Classic/PCR/Liquid Cultures/DNa Clean and Concentrator Prep
Miniprepped the three MLC PCR Constructs. Amplified the BBa_J23119/BBa_K1033931, BBa_J23119/BBa_K1033929, BBa_J23119/BBa_K1073023, BBa_K861171/BBa_K1073023, MLC1/BBa_K1033931, and Tripart PCR constructs with a 32 cycle PCR. Made liquid cultures of pSB1C3 and BBa_B0034 and used the clean kit to prep the PCR products.
September 11th, 2015
Connor
ZR Plasmid Miniprep Classic/Elongated Ligation/Elongated Digestion/Transformation
Miniprepped the pSB1C3 and BBa_B0034 cultures. Digested the PCR constructs, pSB1C3 and BBa_B0034 stocks and incubated for two hours. Ligated the PCR constructs together with the backbones and incubated for 3 hours. Transformed and incubated overnight.
September 12th, 2015
Connor
Liquid Cultures
Prepared liquid cultures of the BBa_J23119/BBa_K1073023, BBa_J23119/BBa_K1033929, and BBa_K118011/BBa_K1033929 PCR constructs.
September 13th, 2015
Jerry
Characterized the sensitivity parts by varying the amount of glucose concentrations in liquid cultures and monitoring their growth and changes in absorbance over time and with based on glucose concentrations.
Connor
ZR Plasmid Miniprep Classic
Miniprepped pSB1C3 liquid cultures and the BBa_K861171/BBa_K1073023 stocks. Nanodropped and stored.
September 14th, 2015
Connor
Elongated Digestion/Elongated Ligation/Transformation
Digested three pSB1C3 stocks and one BBa_B0034 stock along with 500 ng of purified PCR constructs. Incubated for three hours. Ligated together to insert the constructs into the backbone and incubated for 3 hours. Transformed and incubated overnight.
September 15th, 2015
Connor
Liquid Culture
Made liquid cultures of all of the viable PCR constructs, excluding the MLC1 construct and the full constructs.
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