IGEM:UNAM-Genomics-Mexico/2009/Notebook/iGEM 2011 UNAM-Genomics Mexico/2011/05/31: Difference between revisions

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(Autocreate 2011/05/31 Entry for IGEM:UNAM-Genomics-Mexico/2009/Notebook/iGEM_2011_UNAM-Genomics_Mexico)
 
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==Entry title==
==PBBRMCS-5 DNA Plasmid extraction and Digestion==
* Insert your content here.


PBBRMCS-5 was extracted using alcaline lysis and 15μL Plasmid DNA digested with XBA I and Hind III


====PBBRMCS-5 Plasmid DNA extracttion====
We used Miguel Angel Ramirez's Method without the steps 11, and 13 through 20.
=====Gel Electrophoresis=====
The gel was made with 100 mL of TAE 1X and 1 gr. of Agarose, to have a gel with 1% agarose.
* 1st Lane -> 5 μL of Ladder (Fermentas)
* 2nd Lane -> 5 μL PBBRMCS-5 DNA Plasmid, 2 μL loading DNA dye. (1)
* 3rd Lane -> 5 μL PBBRMCS-5 DNA Plasmid, 2 μL loading DNA dye. (2)
* 4th Lane -> 5 μL PBBRMCS-5 DNA Plasmid, 2 μL loading DNA dye. (3)
* 5th Lane -> 5 μL PBBRMCS-5 DNA Plasmid, 2 μL loading DNA dye. (4)
* 6th Lane -> 2 μL of Loading DNA Dye.
* 7th Lane -> 5 μL of Ladder (Fermentas)
====PBBRMCS-5 Plasmid Digestion====
* 2.5 μL of H20
* 2.5 μL of NEBuffer 2 10X
* 1.25 μL of XBA I
* 1.25 μL of Hind III
* 2.5 μL of BSA
* 15 μL of PBBRMCS-5 Plasmid DNA
For a total reaction of 25 μL.
Leave all night at 37ºC in agitation.


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PBBRMCS-5 DNA Plasmid extraction and Digestion

PBBRMCS-5 was extracted using alcaline lysis and 15μL Plasmid DNA digested with XBA I and Hind III

PBBRMCS-5 Plasmid DNA extracttion

We used Miguel Angel Ramirez's Method without the steps 11, and 13 through 20.

Gel Electrophoresis

The gel was made with 100 mL of TAE 1X and 1 gr. of Agarose, to have a gel with 1% agarose.

  • 1st Lane -> 5 μL of Ladder (Fermentas)
  • 2nd Lane -> 5 μL PBBRMCS-5 DNA Plasmid, 2 μL loading DNA dye. (1)
  • 3rd Lane -> 5 μL PBBRMCS-5 DNA Plasmid, 2 μL loading DNA dye. (2)
  • 4th Lane -> 5 μL PBBRMCS-5 DNA Plasmid, 2 μL loading DNA dye. (3)
  • 5th Lane -> 5 μL PBBRMCS-5 DNA Plasmid, 2 μL loading DNA dye. (4)
  • 6th Lane -> 2 μL of Loading DNA Dye.
  • 7th Lane -> 5 μL of Ladder (Fermentas)

PBBRMCS-5 Plasmid Digestion

  • 2.5 μL of H20
  • 2.5 μL of NEBuffer 2 10X
  • 1.25 μL of XBA I
  • 1.25 μL of Hind III
  • 2.5 μL of BSA
  • 15 μL of PBBRMCS-5 Plasmid DNA

For a total reaction of 25 μL.

Leave all night at 37ºC in agitation.


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