Team:Arizona State/Notebook/Brady
LuxI Transformation
Made with Benchling
Project: Brady Dennison IGEM
Authors: Brady Dennison
Date: 2016-06-08
Wednesday, 6/8/16
10 ul of water
.5 ul of DNA (LuxI in this case)
30ul of BL21
Incubate on Ice for 10 min
Warm Plates thorughout all of this, and label during incubation
Place total content onto center of plate
Add a few glass beades
Swirl in infinity style
Remove beads (Special container, they are recycled)
Place the plates into 37 degree Celsius Incubator
BraR Transformation
Made with Benchling
Project: Brady Dennison IGEM
Authors: Brady Dennison
Date: 2016-06-28
Tuesday, 6/28
10 ul of water
.5 ul of DNA (BRAr in this case)
30ul of BL21
Incubate on Ice for 10 min
Warm Plates thorughout all of this, and label during incubation
Place total content onto center of plate
Add a few glass beads
Swirl in infinity style
Remove beads (Special container, they are recycled)
Place the plates into 37 degree Celsius Incubator
BraR Culture
Made with Benchling
Project: Brady Dennison IGEM
Authors: Brady Dennison
Date: 2016-06-29
Wednesday, 6/29
●
Took out transformed plate, selected 4 colonies and labeled them (1-4).
●
Created 4 15 mL tubes with 4 mL of LB Amp, and labeled 1-4.
●
Took a plate from the fridge, and divided it into 4 equal parts, and labeled 1-4.
●
Using a pipette tip, took some bacteria from seleced colonies, then streaked onto plate, then added the pipette tip to the appropriately labeled 15mL tube.
●
Placed tubes into shaking incubator overnight, placed transformed plate into fridge, and placed the newly streaked plate into 37 degree incubator.
BraR Plasmid Extraction/Sequencing
Made with Benchling
Project: Brady Dennison IGEM
Authors: Brady Dennison
Date: 2016-06-30
Thursday, 6/30
Readings:
Nanodrop Readings:
A | B | |
1 | Sample | Concentration (ng/ul) |
2 | 1 | 235 |
3 | 2 | 152 |
4 | 3 | 182.53 |
5 | 4 | 305.28 |
Table1
Sequencing:
A | B | C | D | E | |
1 | What is added | Concentration | Amount Added | ||
2 | Sample 4 | 305.28 ng/uL | .6 uL | ||
3 | Primer (Forward and Reverse) | 10 umol/uL | .5 uL | ||
4 | Water | 5 uL | |||
5 | Total | 6.1 uL |
Table2
Sequencing Results:
LasR NanoDrop/Gel
Made with Benchling
Project: Brady Dennison IGEM
Authors: Brady Dennison
Date: 2016-07-14
Thursday, 7/14
Nano drop:
A | B | C | |
1 | LasR 1 | 0.44 | 0.413 |
2 | 0.232 | 0.218 | |
3 | 1.897 | 1.89 | |
4 | 439.598 | 412.76 | |
5 | LasR 2 | 0.361 | 0.356 |
6 | 0.19 | 0.188 | |
7 | 1.896 | 1.897 | |
8 | 361.007 | 356.49 | |
9 | LasR 3 | 0.379 | 0.371 |
10 | 0.201 | 0.196 | |
11 | 1.89 | 1.89 | |
12 | 379.256 | 370.841 |
Table1
Enzyme Digest Table:
A | B | |
1 | What Did I Put In? | |
2 | Water | 8uL |
3 | EcoRI | 1uL |
4 | KpnI | 1uL |
5 | DNA | 2/3/3 uL (Las 1-3) |
6 | Buffer (Green 10x) | 2 uL |
Table2
BraR, AubI, AubR
Made with Benchling
Project: Brady Dennison IGEM
Authors: Brady Dennison
Date: 2016-07-26
Tuesday, 7/26
A | B | C | D | E | F | G | H | I | J | |
1 | Concentration (ug/uL) | Amount Added (DNA) uL | Amount Added (Primer) uL | Amount Added (Water) uL | ||||||
2 | AubI - 1 | 383.1 | 0.5 | 0.5 | 5 | |||||
3 | AubI - 2 | 88.1 | 2 | 0.5 | 3.5 | |||||
4 | AubI - 3 | 117.8 | 1.5 | 0.5 | 4 | |||||
5 | AubR - 2 | 209.3 | 0.9 | 0.5 | 4.6 | |||||
6 | AubR - 3 | 193.7 | 1 | 0.5 | 4.5 | |||||
7 | BraR - 1 | 290.614 | 0.6 | 0.5 | 4.9 | |||||
8 | BraR - 2 | 533.824 | 0.3 | 0.5 | 5.2 | |||||
9 | BraR - 3 | 387.41 | 0.5 | 0.5 | 5 | |||||
10 |
Table1
Untitled
Made with Benchling
Project: Brady Dennison IGEM
Authors: Brady Dennison
Date: 2016-09-08
Thursday, 9/8
Untitled
Made with Benchling
Project: Brady Dennison IGEM
Authors: Brady Dennison
Date: 2016-09-08
Thursday, 9/8
A | B | C | |
1 | 10 ng AubR | Negative Cntrl | Positive Cntrl |
2 | 5 uL 10x reaction Buffers | 10uL 5X phusion buffer | 10uL 5X phusion buffer |
3 | 2 ul AubR Template | 0 uL template | 2uL pWhitescript 4.5kb control |
4 | 1 uL primer 1 | 1 uL primer 1 | 1 uL cntrl primer 1 |
5 | 1 uL primer 2 | 1 uL primer 2 | 1 uL cntrl primer 2 |
6 | 1uL of dNTPs | 1uL of dNTPs | 1uL of dNTPs |
7 | 32 uL ddH2O (to 50uL) | 36 uL ddH2O (to 50uL) | 34 uL ddH2O (to 50uL) |
8 | 1uL of Phusion pol | 1uL of Phusion pol | 1uL of Phusion pol |
Table1
Induction Experiment
Made with Benchling
Project: Brady Dennison IGEM
Authors: Brady Dennison
Date: 2016-09-29
Thursday, 9/29
Set up 8 10 mL round bottom culture tubes
- 4 tubes containing inducers
- 4 tubes containing controls/receivers
