Team:Groningen/protocols/Ligation

From 2013.igem.org

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<br>All the reagents are added following the order listed in the table above. When the reaction mix is prepared, the content is mixed and incubated at room temperature (~22&deg;C) for 20 min.
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<br>All the reagents are added following the order listed in the table above.
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<br>After the reaction is ready mix the content of the tube and spin it down.
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<br>The tubes are incubated for at room temperature (~22&deg;C) for 1h.
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<br>Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf
<br>Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf

Revision as of 15:33, 27 July 2013

Ligation

Materials:

  • MQ water
  • 1.5ml tubes
  • Ligation buffer
  • Ligase
  • Insert
  • Vector

Reaction:

Component 20µl Final concentration
MilliQ water up to 20µl
10 ligation buffer 2µl 1x
Vector xµl 20-100ng
Insert xµl 3:1 molar ratio over vector
T4 DNA ligase 5U/µl 2µl 0.5U/µl

All the reagents are added following the order listed in the table above. When the reaction mix is prepared, the content is mixed and incubated at room temperature (~22°C) for 20 min.
Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf