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Team:Alberta/Notebook/protocols/LB
From 2010.igem.org
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{{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}} | ||
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| - | == | + | ==Making LB Plates== |
| - | === | + | ===LB Agar (To Make 1L):=== |
| - | + | ||
| - | LB Agar (To Make 1L): | + | |
*Before you start: | *Before you start: | ||
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** DON`T put tubes of antibiotics back into freezer. | ** DON`T put tubes of antibiotics back into freezer. | ||
* Add the following to 800 ml MilliQ H20: | * Add the following to 800 ml MilliQ H20: | ||
| - | + | ::Bacto-tryptone || 10g | |
| - | + | ::Yeast extract || 5g | |
| - | + | ::NaCl || 10g | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
* Adjust pH to 7.5 with NaOH (Optional). | * Adjust pH to 7.5 with NaOH (Optional). | ||
* Add 15g agar. | * Add 15g agar. | ||
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* Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder. | * Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder. | ||
* Pour plates, making sure they are color coded with the type of antibiotic that you used. | * Pour plates, making sure they are color coded with the type of antibiotic that you used. | ||
| - | |||
==Making LB Broth:== | ==Making LB Broth:== | ||
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{{Team:Alberta/endMainContent}} | {{Team:Alberta/endMainContent}} | ||
Latest revision as of 02:21, 27 October 2010
Making LB Plates
LB Agar (To Make 1L):
- Before you start:
- Use AGAR not agarose!
- Use an Erlenmeyer flask that is twice the volume of your solution (avoid spillage in autoclave).
- Keep your flask on a stir plate with a stir rod.
- Put Aluminum foil on the top of the flask and put autoclave tape on.
- DON`T let it solidify after autoclaving by keeping it on a stir plate while it cools.
- DON`T put tubes of antibiotics back into freezer.
- Add the following to 800 ml MilliQ H20:
- Bacto-tryptone || 10g
- Yeast extract || 5g
- NaCl || 10g
- Adjust pH to 7.5 with NaOH (Optional).
- Add 15g agar.
- Melt agar into solution in the microwave (Optional).
- Adjust volume to 1L with MilliQ H20
- Autoclave on the liquid cycle (no drying time) (aka LIQ20) for 30 minutes.
- Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder.
- Pour plates, making sure they are color coded with the type of antibiotic that you used.
Making LB Broth:
- All of the above WITHOUT AGAR.
