Team:IPN-UNAM-Mexico/Notebook/August 10 2009

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August 10th, 2009

Calculus for Mix; plasmid linearization.

For 20 reactions.

 

 
Mix
For every Eppendorf tube
For 20 reactions

PlasmidicDNA

3 μl

60 μl

Buffer 2

1 μl

20 μl

BSA

0.5μl

10 μl

EcoRI

0.5 μl / 5 μll

10 μl / 100 μl

H2O

5 μl / 10 μl   vf

100 μl / 200 μl   vf

 

 

 

 

 

 

Electrophoresis gel load:

  • 3 μl of colorant
  • 10 μl of digestion
  • 3 μl of molecular weight marker

 

Load order.

Well     Plasmid            Well             Plasmid

2          16,2”               13            3.3
3          4:23 L2            14            22.1
4          15:3 L2            15            19.2*
5          18.2*               16            24.2”
6          2.3

Well                             Well
1          1kb 2 μl         12            2.3
2          16.2”             13            14.2
3          4:23 L2          14            3.3
4          15:3 L2          15            22.1
5          18.2              15            19.1*
6          15.13

 

Horizontal Gene Transfer (HGT).
Acording to the model proposed to Arturo, we made a search for the candidate bioparts to make their elusion and transformation, which got in the next way:

2 Constructions:


Construction 1:
Biopart BBa_K145201

J23116 + B0034 + C0040 + B0010 + B0012

Construction 2:
The first option for the construction 2 is made up from two devices: BBa_J01101 and Bba_I763011. The second one consists in asking Boston for the construction BBa_I763027. We will tell Arturo about this posibility.
First option:
Device 1.
B0040 + B0034 + C0051 + B0010 + B0012
Device 2.
B0051 + B0034 + J04031

 

LOCATION

 

Plate

Well

Plasmid

Device 1 (piece1)

2009 kitplate1

11H

PSB1A2

Device 2 (piece 2)

2009 kitplate1

15L

PSB1A2

 

 

 

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