Team:TU Delft/23 June 2010 content

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{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
|'''#'''
|'''#'''
-
|'''Ligation reaction'''
+
|'''BioBrick'''
 +
|'''Fragment 1'''
 +
|'''Fragment 2'''
 +
|'''Recipient vector'''
|-
|-
|1
|1
-
|3 μL I10341 + 4 μL J1100 + 1.5 μL pSB3C5
+
|
 +
|4 μL μL ‘E–J1100–S’
 +
|3 μL μL ‘X–I10341–P’
 +
|1.5 μL ‘E–linear pSB1T3–P’
|-
|-
|2
|2
-
|3 μL I10341 + 4 μL J1101 + 1.5 μL pSB3C5
+
|
 +
|4 μL μL ‘E–J1101–S’
 +
|3 μL μL ‘X–I10341–P’
 +
|1.5 μL ‘E–linear pSB1T3–P’
|-
|-
|3
|3
-
|3 μL I10341 + 4 μL J1107 + 1.5 μL pSB3C5
+
|
 +
|4 μL μL ‘E–J1107–S’
 +
|3 μL μL ‘X–I10341–P’
 +
|1.5 μL ‘E–linear pSB1T3–P’
|-
|-
|4
|4
-
|3 μL I10341 + 4 μL J1117 + 1.5 μL pSB3C5
+
|
 +
|4 μL μL ‘E–J1117–S’
 +
|3 μL μL ‘X–I10341–P’
 +
|1.5 μL ‘E–linear pSB1T3–P’
|-
|-
|5
|5
-
|3 μL I10341 + 4 μL J1127 + 1.5 μL pSB3C5
+
|
 +
|4 μL μL ‘E–J1127–S’
 +
|3 μL μL ‘X–I10341–P’
 +
|1.5 μL ‘E-linear pSB1T3–P’
|-
|-
|6
|6
-
|4 μL J1100 + 1.5 μL pSB3C5 (negative control)
+
|negative control
 +
|'''-'''
 +
|3 μL μL ‘X–I10341–P’
 +
|1.5 μL ‘E–linear pSB1T3–P’
|}
|}
We adhered to the 3:1 insert:plasmid ratio when determining the volumes of DNA added.
We adhered to the 3:1 insert:plasmid ratio when determining the volumes of DNA added.
The mixtures were incubated overnight at 16 °C.
The mixtures were incubated overnight at 16 °C.

Revision as of 16:54, 31 July 2010

Lab work

Characterization of Anderson RBS sequences

Ligations were performed using the overnight digested BioBricks. The following ligation reactions was performed:

# BioBrick Fragment 1 Fragment 2 Recipient vector
1 4 μL μL ‘E–J1100–S’ 3 μL μL ‘X–I10341–P’ 1.5 μL ‘E–linear pSB1T3–P’
2 4 μL μL ‘E–J1101–S’ 3 μL μL ‘X–I10341–P’ 1.5 μL ‘E–linear pSB1T3–P’
3 4 μL μL ‘E–J1107–S’ 3 μL μL ‘X–I10341–P’ 1.5 μL ‘E–linear pSB1T3–P’
4 4 μL μL ‘E–J1117–S’ 3 μL μL ‘X–I10341–P’ 1.5 μL ‘E–linear pSB1T3–P’
5 4 μL μL ‘E–J1127–S’ 3 μL μL ‘X–I10341–P’ 1.5 μL ‘E-linear pSB1T3–P’
6 negative control - 3 μL μL ‘X–I10341–P’ 1.5 μL ‘E–linear pSB1T3–P’

We adhered to the 3:1 insert:plasmid ratio when determining the volumes of DNA added. The mixtures were incubated overnight at 16 °C.

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