Team:Alberta/Notebook/protocols/ligation

From 2010.igem.org

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==General Protocols==
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/invitro_biobyte_assembly | In Vitro BioByte Assembly]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/LB | LB Plates and Broth]]
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*[[Team:Alberta/Notebook/protocols/transformations |Transformations]]
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*[[Team:Alberta/Notebook/protocols/overnight |5mL Overnight ]]
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*[[Team:Alberta/Notebook/protocols/glycerol | Glycerol Stock ]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/miniprep | Plasmid Miniprep ]]
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*[[Team:Alberta/Notebook/protocols/digest | Restriction Digest ]]
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*[[Team:Alberta/Notebook/protocols/vector_dephos | Vector Dephosphorylation]]
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*[[Team:Alberta/Notebook/protocols/ligation | Ligation ]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/agarose_gel | Agarose Gel Electrophoresis ]]
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*[[Team:Alberta/Notebook/protocols/gel_extraction | Gel Extraction ]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/pcr | PCR]]
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*[[Team:Alberta/Notebook/protocols/colony_pcr | Colony PCR ]]
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*[[Team:Alberta/Notebook/protocols/pcr_purification | PCR Purification ]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/labelling | Sample Labelling Conventions]]
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*[[Team:Alberta/Notebook/protocols/sequencing | Fluorescent Sequencing Reaction]]
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*[[Team:Alberta/Notebook/protocols/primer_design | Primer Design]]
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-----------------------------
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Latest revision as of 02:29, 27 October 2010

TEAM ALBERTA


Ligation

Reagents:

  • Vector
  • Insert
  • MilliQ buffer
  • 5X ligase buffer
  • T4 DNA ligase
    • [http://tools.invitrogen.com/content/sfs/manuals/t4dnaligase_5U_man.pdf]


Procedure:

  • Volumes of vector and insert may vary, but a typical 20 ul reaction might be:
MilliQ water 11 ul
5x ligase buffer 4 ul
Vector (200 ng/ul) 1 ul
Insert (200 ng/ul) 3 ul
Ligase 1 ul
  • Incubate at R/T for 1 hour or overnight at 12-16oC.


Retrieved from "http://2010.igem.org/Team:Alberta/Notebook/protocols/ligation"