Lidstrom:Protocols: Difference between revisions

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*[[Lidstrom:Health & Safety|Health & Safety]]
*[[Lidstrom:Health & Safety|Health & Safety]]
*[[Lidstrom:Finding Chemicals|Finding Chemicals]]
*[[Lidstrom:Finding Chemicals|Finding Chemicals]]
*[[Lidstrom:Dishwasher |Dishwasher]]
*[[Lidstrom:Autoclave |Autoclave]]
*[[Lidstrom:Autoclave |Autoclave]]
*[[Lidstrom:Sterile Technique | Sterile technique]]
*[[Lidstrom:Sterile Technique | Sterile technique]]
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==Cell Culture:==
==Cell Culture:==
*[[Lidstrom:Tube Spec|Tube Spec]]
*[[Lidstrom:Tube Spec|Tube Spec]]
*[[Lidstrom:M9|M9 recipe]]
*[[Lidstrom:TB|Terrific Broth recipe]]
*[[Lidstrom: Bioscreen|Bioscreen Setup]]
*[[Lidstrom: EZ amino acid stock recipe|EZ amino acid stock recipe]]


==DNA/RNA:==
==DNA/RNA:==


===DNA===
===DNA===
====Isolation ====
*[[Lidstrom:QIAquick Gel Extraction Kit Protocol|QIAquick Gel Extraction Kit Protocol]]
*[[Lidstrom:Genomic DNA extraction|Genomic DNA extraction]]
====Manipulation====
*[[Lidstrom:Building with DNA|Intro: Building with DNA (in progress)]]
*[[Lidstrom:Building with DNA|Intro: Building with DNA (in progress)]]
*[[Lidstrom:Colony PCR| Colony PCR]]
*[[Lidstrom:Digestion with Restriction Enzymes|Digestion with Restriction Enzymes]]
*[[Lidstrom:Digestion with Restriction Enzymes|Digestion with Restriction Enzymes]]
*[[Lidstrom:Diluting Primers | Diluting Primers]]
*[[Lidstrom:Gibson Assembly| Gibson Assembly (In progress)]]
*[[Lidstrom:Gibson Assembly| Gibson Assembly (In progress)]]
*[[Lidstrom:Ligation |Ligation (in progress)]]
*[[Lidstrom:Ligation |Ligation (in progress)]]
*[[Lidstrom:Loading Dye Recipe|Loading Dye Recipe]]
*[[Lidstrom:Miniprep |Miniprep (in progress)]]
*[[Lidstrom:Oligo Orders | Oligo Orders (in progress)]]
*[[Lidstrom:Overlap Extension PCR| Overlap Extension PCR (in progress)]]
*[[Lidstrom:Overlap Extension PCR| Overlap Extension PCR (in progress)]]
*[[Qiagen_Buffers| Qiagen Buffer Recipes]]
=====Fancy Manipulation=====
*[[Lidstrom:Site-Directed Mutagenesis|Site-Directed Mutagenesis (in progress)]]
*[[Lidstrom:λ-Red Mediated Gene Knockouts|λ-Red Mediated Gene Knockouts (in progress)]]
*[[Dorman:P1_phage_%28lysogenic%29|Phage Lysate Prep]]
*[[Lidstrom:P1 Phage Transduction|P1 Phage Transduction (in progress)]]
====Obervation====
*[[Lidstrom:Colony PCR| Colony PCR]]
*[[Lidstrom: Agarose Gel Electrophoresis|Agarose Gel Electrophoresis]]
*[[Lidstrom:Sequencing with GeneWiz|Sequencing with GeneWiz]]
====Misc.====
*[[Lidstrom:PCR| PCR (in progress)]]
*[[Lidstrom:PCR| PCR (in progress)]]
*[[Lidstrom:Loading Dye Recipe|Loading Dye Recipe]]
*[[Lidstrom:Purifying DNA|Purifying DNA (in progress)]]
*[[Lidstrom:Purifying DNA|Purifying DNA (in progress)]]
*[[Lidstrom:Sequencing with GeneWiz|Sequencing with GeneWiz]]
*[[Lidstrom:Miniprep |Miniprep (in progress)]]
*[[Lidstrom:Site-Directed Mutagenesis|Site-Directed Mutagenesis (in progress)]]
*[[Lidstrom:Oligo Orders | Oligo Orders]]
*[[Lidstrom:Transformation| Transformation (in progress)]]
*[[Lidstrom:Diluting Primers | Diluting Primers]]


===RNA===
==Protein==
*[[Lidstrom: SDS-PAGE | SDS-PAGE Protein Gels]]


==E. Coli==
==E. Coli==
*[[Lidstrom:E. Coli Vector Compatibility| E. Coli Vector Compatibility  (In Progress)]]
*[[Lidstrom:Competent Cell Preparation|Competent Cell Preparation (in progress)]]
*[[Lidstrom:Competent Cell Preparation|Competent Cell Preparation (in progress)]]
*[[Lidstrom:Chemical Transformation|Chemical Transformation]]
*[[Lidstrom:Electroporation|Electroporation]]
*[[Lidstrom:Competent Cells|Competent Cell Lines]]
==Enzyme Assays==
===General===
*[[Lidstrom: Plate Reader| Plate Reader Setup]]
===in vitro pathway assays (CO<sub>2</sub> project)===
*[[Lidstrom:in vitro Pathway Assay: Cell Prep| Cell Prep (in progress)]]
*[[Lidstrom:in vitro Pathway Assay: Crude Extract Prep| Crude Extract Prep (in progress)]]
*[[Lidstrom:in vitro Pathway Assay: Sample Analysis via UPLC-MS/MS | Sample Analysis via UPLC-MS/MS (in progress)]]


==Electrocompetent Cells==
== Metabolomics ==
*You can make your own electro-competent cells for electroporation.
=== CO2 project ===
**From Amada's past mentor in undergrad:
*[[Lidstrom:Measuring 13C Incorporation Into Protein - CO2 Project|Measuring 13C Incorporation Into Protein - CO2 Project]]
***Grow the cells overnight
** [[Lidstrom:13C Incorporation Into Protein - Data Analysis]]
***Inoculate from this the next morning: generally use 200 uL/50 mL
***Let grow for about 3-4 h (OD 0.4-0.6 or so),  then centrifuge and wash 3 times with 10% glycerol (everything on ice).
***Resuspend the pellet in a small volume (Example, if I do a 5 ml culture I try to resuspend the final pellet in no more than 0.2 or 0.3 ml of 10% glycerol). Then split in 1.5 ml centrifuge tubes (0.1 ml in each), freeze on dry ice-isopropanol and store at -80C.
****Note: you can dilute your re-suspension, measure OD, and calculate the cell concentration if you care.  See [[Electrocompetent_Cells| this page]].  Would allow you to calculate efficiency.

Revision as of 19:25, 10 February 2013

Back home: Lidstrom

General Lab:

Cell Culture:

DNA/RNA:

DNA

Isolation

Manipulation

Fancy Manipulation

Obervation

Misc.

Protein

E. Coli

Enzyme Assays

General

in vitro pathway assays (CO2 project)

Metabolomics

CO2 project