"
Team:Alberta/Notebook/protocols/ligation
From 2010.igem.org
(Difference between revisions)
(New page: {{Team:Alberta/Head}} {{Team:Alberta/navbar|project=selected}} {{Team:Alberta/beginLeftSideBar}} {{Team:Alberta/endLeftSideBar}} {{Team:Alberta/beginRightSideBar}} {{Team:Alberta/endRig...) |
|||
| (2 intermediate revisions not shown) | |||
| Line 1: | Line 1: | ||
{{Team:Alberta/Head}} | {{Team:Alberta/Head}} | ||
| - | {{Team:Alberta/navbar| | + | {{Team:Alberta/navbar|notebook=selected}} |
| + | |||
| + | {{Team:Alberta/beginLeftSideBar|toc=NO}} | ||
| + | |||
| + | |||
| + | ==General Protocols== | ||
| + | ----------------------------- | ||
| + | *[[Team:Alberta/Notebook/protocols/invitro_biobyte_assembly | In Vitro BioByte Assembly]] | ||
| + | ----------------------------- | ||
| + | *[[Team:Alberta/Notebook/protocols/LB | LB Plates and Broth]] | ||
| + | |||
| + | *[[Team:Alberta/Notebook/protocols/transformations |Transformations]] | ||
| + | |||
| + | *[[Team:Alberta/Notebook/protocols/overnight |5mL Overnight ]] | ||
| + | |||
| + | *[[Team:Alberta/Notebook/protocols/glycerol | Glycerol Stock ]] | ||
| + | ----------------------------- | ||
| + | *[[Team:Alberta/Notebook/protocols/miniprep | Plasmid Miniprep ]] | ||
| + | |||
| + | *[[Team:Alberta/Notebook/protocols/digest | Restriction Digest ]] | ||
| + | |||
| + | *[[Team:Alberta/Notebook/protocols/vector_dephos | Vector Dephosphorylation]] | ||
| + | |||
| + | *[[Team:Alberta/Notebook/protocols/ligation | Ligation ]] | ||
| + | ----------------------------- | ||
| + | *[[Team:Alberta/Notebook/protocols/agarose_gel | Agarose Gel Electrophoresis ]] | ||
| + | |||
| + | *[[Team:Alberta/Notebook/protocols/gel_extraction | Gel Extraction ]] | ||
| + | ----------------------------- | ||
| + | *[[Team:Alberta/Notebook/protocols/pcr | PCR]] | ||
| + | |||
| + | *[[Team:Alberta/Notebook/protocols/colony_pcr | Colony PCR ]] | ||
| + | |||
| + | *[[Team:Alberta/Notebook/protocols/pcr_purification | PCR Purification ]] | ||
| + | ----------------------------- | ||
| + | *[[Team:Alberta/Notebook/protocols/labelling | Sample Labelling Conventions]] | ||
| + | |||
| + | *[[Team:Alberta/Notebook/protocols/sequencing | Fluorescent Sequencing Reaction]] | ||
| + | |||
| + | *[[Team:Alberta/Notebook/protocols/primer_design | Primer Design]] | ||
| + | ----------------------------- | ||
| + | |||
| + | |||
| + | {{Team:Alberta/endMainContent}} | ||
| + | |||
| - | |||
{{Team:Alberta/endLeftSideBar}} | {{Team:Alberta/endLeftSideBar}} | ||
| Line 11: | Line 54: | ||
{{Team:Alberta/beginMainContent}} | {{Team:Alberta/beginMainContent}} | ||
| - | + | ==Ligation== | |
| - | Reagents: | + | ===Reagents:=== |
* Vector | * Vector | ||
| Line 24: | Line 67: | ||
| - | Procedure: | + | ===Procedure:=== |
*Volumes of vector and insert may vary, but a typical 20 ul reaction might be: | *Volumes of vector and insert may vary, but a typical 20 ul reaction might be: | ||
| Line 41: | Line 84: | ||
| - | |||
{{Team:Alberta/endMainContent}} | {{Team:Alberta/endMainContent}} | ||
Latest revision as of 02:29, 27 October 2010
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.
