Team:BU Wellesley Software/Notebook/ShannonNotebook

From 2011.igem.org

(Difference between revisions)
(→6/06/2011-6/10/2011)
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* plasmid preps were made from the previous week’s transformations.  
* plasmid preps were made from the previous week’s transformations.  
** this was done to improve the Nanodrop quantification values.  
** this was done to improve the Nanodrop quantification values.  
-
**plasmid preps didn’t grow properly (only UV plasmid).  
+
** plasmid preps didn’t grow properly (only UV plasmid).  
* an issue with the plates was discovered. All previous cultures were done on Ampicillian plates, and most were not growing well. We switched to a different antibiotic (Kanamycin) and all proceeding cultures grew well.  
* an issue with the plates was discovered. All previous cultures were done on Ampicillian plates, and most were not growing well. We switched to a different antibiotic (Kanamycin) and all proceeding cultures grew well.  
* new Ampicillian plates were made.  
* new Ampicillian plates were made.  
Line 22: Line 22:
** two seemed to look pinkish, especially towards the center of each colony.  
** two seemed to look pinkish, especially towards the center of each colony.  
** successful transformations were then used to produce more plasmid preps.
** successful transformations were then used to produce more plasmid preps.
 +
 +
 +
== 6/20/2011-6/24/2011  ==
 +
* this week plasmid preps were done on the following: Bba_B0015.1, Bba_B0015.2, Bba_B0015.3, Bba_B0015.4, Bba_R0040.1, Bba_R0040.2, Bba_J52028.1, Bba_J52028.2, Bba_J52028.3,and Bba_J52028.4.
 +
* Nanodrop was also conducted. Chart below shows values from 6/23/2011.
 +
 +
{| class="wikitable"
 +
|-
 +
! Sample #
 +
! ng/uL
 +
! 260/280
 +
! 260/230
 +
|-
 +
| Bba_E0240
 +
| 2.70
 +
| 3.54
 +
| 0.01
 +
|-
 +
| Bba_E0430
 +
| 1.5
 +
| -32.2
 +
| 0.01
 +
|-
 +
| Bba_B0015
 +
| 4.5
 +
| 2.69
 +
| 0.03
 +
|-
 +
| Bba_I14033.1
 +
| 3.7
 +
| 4.41
 +
| 0.02
 +
|-
 +
| Bba_I14033.2
 +
| 6.3
 +
| 1.81
 +
| 0.04
 +
|-
 +
| Bba_I13453.1
 +
| 2.3
 +
| 3.71
 +
| 0.01
 +
|-
 +
| Bba_I13453.2
 +
| 1.8
 +
| 29.53
 +
| 0.02
 +
|-
 +
| Bba_I13453.3
 +
| 1.9
 +
| 1.93
 +
| 0.01
 +
|}
 +
 +
* Ligation was also performed. I tried ligating BFP (Bba_K156010) with terminator (Bba_B0015.2.2). Transformation was done of this ligation later that night. The following morning the transformation plates did not have any colonies on the
 +
* Minipreps were conducted on 6/23/2011’s plasmid prep and s Nanodrop quantification was then performed. Results are shown below in the following chart:
 +
 +
{| class="wikitable"
 +
|-
 +
! Sample #
 +
! ng/uL
 +
! 260/280
 +
! 260/230
 +
|-
 +
| Bba_B0015.1,
 +
| 24.5
 +
| 1.80
 +
| 1.78
 +
|-
 +
| Bba_B0015.2
 +
| 20.8
 +
| 1.72
 +
| 1.74
 +
|-
 +
| Bba_B0015.3
 +
| 25.0
 +
| 1.74
 +
| 1.60
 +
|-
 +
| Bba_B0015.4
 +
| 23.2
 +
| 1.86
 +
| 1.95
 +
|-
 +
| Bba_R0040.1
 +
| 20.4
 +
| 1.75
 +
| 1.46
 +
|-
 +
| Bba_R0040.2
 +
| 14.5
 +
| 1.62
 +
| 0.80
 +
|-
 +
| Bba_J52028.1
 +
| 42.0
 +
| 1.89
 +
| 1.98
 +
|-
 +
| Bba_J52028.2
 +
| 54.1
 +
| 1.77
 +
| 1.35
 +
|-
 +
| Bba_J52028.3
 +
| 42.5
 +
| 1.80
 +
| 2.12
 +
|-
 +
| Bba_J52028.4
 +
| 41.5
 +
| 1.91
 +
| 2.03
 +
|}

Revision as of 15:37, 13 July 2011

6/06/2011-6/10/2011

  • included a boot camp.
  • different topics were discussed (i.e. biobricks, gene regulation, differences between eukaryotes and prokaryotes).
  • boot camp also included a computer science component (focused on Clotho).
  • miniprepes were done on previously transformed BFP2. After minipreps, the Nanodrop was used to quantify the DNA. Minipreps did not have high DNA concentration, but were run on a gel (to show presence of DNA). Most lanes appeared empty (bands of DNA were not visible).
  • transformations of Pbad was conducted.
    • The following day, transformations appeared to be contaminated. They looked significantly different than usual, but were still were used to produce plasmid preps.
    • To ensure that E.coli was present gram staining was performed. Staining showed that no E.coli was present in the cultures. Minipreps and Nanodrop were used to quantify the DNA from the plasmid preps.
      • The results from the Miniprep/Nanodrop varied (some plasmid preps had a decent DNA concentration, others didn’t). Even though yields were not great the samples were run on a gel. However no bands were visible.


6/13/2011-6/17/2011

  • plasmid preps were made from the previous week’s transformations.
    • this was done to improve the Nanodrop quantification values.
    • plasmid preps didn’t grow properly (only UV plasmid).
  • an issue with the plates was discovered. All previous cultures were done on Ampicillian plates, and most were not growing well. We switched to a different antibiotic (Kanamycin) and all proceeding cultures grew well.
  • new Ampicillian plates were made.
  • a side project involved ligating GFP (Bba_J52625) and terminator (Bba_B0015) together for future use.
  • transformations were completed (involved transforming seven different biobrick parts).
    • two seemed to look pinkish, especially towards the center of each colony.
    • successful transformations were then used to produce more plasmid preps.


6/20/2011-6/24/2011

  • this week plasmid preps were done on the following: Bba_B0015.1, Bba_B0015.2, Bba_B0015.3, Bba_B0015.4, Bba_R0040.1, Bba_R0040.2, Bba_J52028.1, Bba_J52028.2, Bba_J52028.3,and Bba_J52028.4.
  • Nanodrop was also conducted. Chart below shows values from 6/23/2011.
Sample # ng/uL 260/280 260/230
Bba_E0240 2.70 3.54 0.01
Bba_E0430 1.5 -32.2 0.01
Bba_B0015 4.5 2.69 0.03
Bba_I14033.1 3.7 4.41 0.02
Bba_I14033.2 6.3 1.81 0.04
Bba_I13453.1 2.3 3.71 0.01
Bba_I13453.2 1.8 29.53 0.02
Bba_I13453.3 1.9 1.93 0.01
  • Ligation was also performed. I tried ligating BFP (Bba_K156010) with terminator (Bba_B0015.2.2). Transformation was done of this ligation later that night. The following morning the transformation plates did not have any colonies on the
  • Minipreps were conducted on 6/23/2011’s plasmid prep and s Nanodrop quantification was then performed. Results are shown below in the following chart:
Sample # ng/uL 260/280 260/230
Bba_B0015.1, 24.5 1.80 1.78
Bba_B0015.2 20.8 1.72 1.74
Bba_B0015.3 25.0 1.74 1.60
Bba_B0015.4 23.2 1.86 1.95
Bba_R0040.1 20.4 1.75 1.46
Bba_R0040.2 14.5 1.62 0.80
Bba_J52028.1 42.0 1.89 1.98
Bba_J52028.2 54.1 1.77 1.35
Bba_J52028.3 42.5 1.80 2.12
Bba_J52028.4 41.5 1.91 2.03
Retrieved from "http://2011.igem.org/Team:BU_Wellesley_Software/Notebook/ShannonNotebook"