Team:Newcastle/Gel electrophoresis

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(Materials)
(Materials)
 
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=Gel electrophoresis=
=Gel electrophoresis=
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[[Image:Newcastle Prep Chr Gel.jpg|250px|right]]
==Materials==
==Materials==
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* 50X TAE buffer
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# 50X TAE buffer
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*# Make up 1 liter of 50X TAE buffer with the following:
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#* Make up 1 liter of 50X TAE buffer with the following:
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*# 242g of TRIS base
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#* 242 g of TRIS base
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*# 57.1 ml of acetic acid
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#* 57.1 ml of acetic acid
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*# 100 ml of 0.5 M of EDTA (pH 8.0)
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#* 100 ml of 0.5 M of EDTA (pH 8.0)
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*# Top up to 1 liter with water
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#* Top up to 1 liter with water
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* SafeView
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# SafeView
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* Agarose
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# Agarose
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* DNA ladder
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# DNA ladder
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* Eppendorf
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# Eppendorf tube
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* Gel making tank
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# Gel making tank
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* Gel running tank
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# Gel running tank
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==Procedures==
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==Procedure==
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# Make up 1% agarose gel (1 g of agarose in 100 ml of TAE buffer) and transfer 60 ml of molten agarose gel into the gel tray with 3 μl of Safeview dye.
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# Make up 1% agarose gel (1 g of agarose per 100 ml of TAE buffer) and using the microwave to dissolve the agarose.
 +
# Allow the molten agarose to cool down before adding 5 μl of Safeview dye.
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# Set up the gel tray containing the gel comb and transfer 60 ml of molten agarose gel into the gel tray (take care not to cover the gel comb).
# Wait for 30 min to allow the gel to harden.
# Wait for 30 min to allow the gel to harden.
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# Transfer harden gel into the gel tank and add TAE buffer until the gel is completely submerge.
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# Transfer harden gel into the gel tank and add TAE buffer until the gel is completely submerged.
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# Depending on the nature of the sample, 3μl of GeneRuler™ 1kb Plus DNA Ladder was used for analysing DNA.  
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# Depending on the nature of the sample, 3 μl of GeneRuler™ 1 kb Plus DNA Ladder is used for analysing DNA.  
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# Loading buffer was then added together with the sample before loading onto the gel matrix.
+
# Loading buffer is then added together with the sample before loading onto the gel matrix.
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# Run gel at 90V until separation is achieved and visualize using the gelDoc.
+
# Run gel at 90V until the desired separation is achieved and visualize using the gelDoc.

Latest revision as of 14:28, 26 October 2010

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Gel electrophoresis

Newcastle Prep Chr Gel.jpg

Materials

  1. 50X TAE buffer
    • Make up 1 liter of 50X TAE buffer with the following:
    • 242 g of TRIS base
    • 57.1 ml of acetic acid
    • 100 ml of 0.5 M of EDTA (pH 8.0)
    • Top up to 1 liter with water
  2. SafeView
  3. Agarose
  4. DNA ladder
  5. Eppendorf tube
  6. Gel making tank
  7. Gel running tank

Procedure

  1. Make up 1% agarose gel (1 g of agarose per 100 ml of TAE buffer) and using the microwave to dissolve the agarose.
  2. Allow the molten agarose to cool down before adding 5 μl of Safeview dye.
  3. Set up the gel tray containing the gel comb and transfer 60 ml of molten agarose gel into the gel tray (take care not to cover the gel comb).
  4. Wait for 30 min to allow the gel to harden.
  5. Transfer harden gel into the gel tank and add TAE buffer until the gel is completely submerged.
  6. Depending on the nature of the sample, 3 μl of GeneRuler™ 1 kb Plus DNA Ladder is used for analysing DNA.
  7. Loading buffer is then added together with the sample before loading onto the gel matrix.
  8. Run gel at 90V until the desired separation is achieved and visualize using the gelDoc.


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