Lidstrom:Protocols

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
Current revision (10:50, 4 September 2014) (view source)
(Fancy Manipulation)
 
(33 intermediate revisions not shown.)
Line 10: Line 10:
*[[Lidstrom:Preparing Freezer Cell Stocks| Preparing Freezer Cell Stocks]]
*[[Lidstrom:Preparing Freezer Cell Stocks| Preparing Freezer Cell Stocks]]
*[[Lidstrom:Pouring Media Plates|Pouring Media Plates]]
*[[Lidstrom:Pouring Media Plates|Pouring Media Plates]]
 +
*[[Lidstrom:Filtration|Filtration]]
==Cell Culture:==
==Cell Culture:==
*[[Lidstrom:Tube Spec|Tube Spec]]
*[[Lidstrom:Tube Spec|Tube Spec]]
*[[Lidstrom: Bioscreen|Bioscreen Setup]]
*[[Lidstrom: Bioscreen|Bioscreen Setup]]
 +
=== Methylotrophs ===
=== Methylotrophs ===
*[[Lidstrom:MM1/MM2/MM3/HY|MM1/MM2/MM3/HY]]
*[[Lidstrom:MM1/MM2/MM3/HY|MM1/MM2/MM3/HY]]
Line 20: Line 22:
*[[Lidstrom:TB|Terrific Broth recipe]]
*[[Lidstrom:TB|Terrific Broth recipe]]
*[[Lidstrom: EZ amino acid stock recipe|EZ amino acid stock recipe]]
*[[Lidstrom: EZ amino acid stock recipe|EZ amino acid stock recipe]]
 +
 +
=== OD measurements ===
 +
* [[Lidstrom:Beckmann DU 640B spectrophotometer|Beckmann DU 640B spectrophotometer]]
 +
*[[Lidstrom:NanoDrop|Nano Drop for Cell Cultures]]
==DNA/RNA:==
==DNA/RNA:==
===DNA===
===DNA===
-
== PCR ==  
+
==== PCR ====  
*[[Lidstrom:Colony PCR| Colony PCR]]
*[[Lidstrom:Colony PCR| Colony PCR]]
*[[Lidstrom:PCR| PCR (in progress)]]
*[[Lidstrom:PCR| PCR (in progress)]]
Line 41: Line 47:
=====Fancy Manipulation=====
=====Fancy Manipulation=====
*[[Lidstrom:Site-Directed Mutagenesis|Site-Directed Mutagenesis (in progress)]]
*[[Lidstrom:Site-Directed Mutagenesis|Site-Directed Mutagenesis (in progress)]]
-
*[[Lidstrom:λ-Red Mediated Gene Knockouts|λ-Red Mediated Gene Knockouts (in progress)]]
+
** [[Lidstrom:QuikChange Site-Directed Mutagenesis|QuikChange Site-Directed Mutagenesis]]
 +
*[[Lidstrom:λ-Red Mediated Gene Knockouts|λ-Red Mediated Gene Knockouts]]
*[[Dorman:P1_phage_%28lysogenic%29|Phage Lysate Prep]]
*[[Dorman:P1_phage_%28lysogenic%29|Phage Lysate Prep]]
*[[Lidstrom:P1 Phage Transduction|P1 Phage Transduction (in progress)]]
*[[Lidstrom:P1 Phage Transduction|P1 Phage Transduction (in progress)]]
Line 56: Line 63:
*[[Lidstrom: Agarose Gel Electrophoresis|Agarose Gel Electrophoresis]]
*[[Lidstrom: Agarose Gel Electrophoresis|Agarose Gel Electrophoresis]]
*[[Lidstrom:Sequencing with GeneWiz|Sequencing with GeneWiz]]
*[[Lidstrom:Sequencing with GeneWiz|Sequencing with GeneWiz]]
 +
 +
====Mutagenesis====
 +
*[[Lidstrom:EMS Mutagenesis|EMS Mutagenesis]]
 +
*[[Lidstrom:UV Mutagenesis|UV Mutagenesis]]
====Misc.====
====Misc.====
Line 63: Line 74:
*[[Lidstrom:Oligo Orders | Oligo Orders]]
*[[Lidstrom:Oligo Orders | Oligo Orders]]
*[[Lidstrom:Diluting Primers | Diluting Primers]]
*[[Lidstrom:Diluting Primers | Diluting Primers]]
 +
*[https://www.neb.com/~/media/NebUs/Files/Brochures/Cloning_Guide_1113.pdf NEB cloning guide]
==Protein==
==Protein==
 +
=== Purification ===
 +
*[[Lidstrom:His-tag Protein Purification|His-tag Protein Purification]]
 +
 +
=== Analytics ===
*[[Lidstrom: SDS-PAGE | SDS-PAGE Protein Gels]]
*[[Lidstrom: SDS-PAGE | SDS-PAGE Protein Gels]]
 +
 +
=== Protein Concentration Determination ===
 +
*[[Lidstrom:Choosing a protein concentration quantification method|Choosing a protein concentration quantification method]]
*[[Lidstrom:BCA assay|BCA assay]] to determine protein concentration
*[[Lidstrom:BCA assay|BCA assay]] to determine protein concentration
 +
*[[Lidstrom:NanoDrop#NanoDrop_for_Proteins|NanoDrop for proteins]]
 +
 +
== Chemicals ==
 +
*[[Lidstrom:Chemical Stability|Chemical Stability]]
 +
* Also, see [[Lidstrom:Solution Stock Info|Solution Stock Info]]
==Buffers==
==Buffers==
*[[Lidstrom:Buffers|Buffers]]
*[[Lidstrom:Buffers|Buffers]]
-
==E. Coli==
+
==E. coli==
-
*[[Lidstrom:E. Coli Vector Compatibility| E. Coli Vector Compatibility  (In Progress)]]
+
*[[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)]]
 +
=== E. coli transformation ===
*[[Lidstrom:Chemical Transformation|Chemical Transformation]]
*[[Lidstrom:Chemical Transformation|Chemical Transformation]]
*[[Lidstrom:Electroporation|Electroporation]]
*[[Lidstrom:Electroporation|Electroporation]]
Line 81: Line 106:
*[[Lidstrom:French Press|French Press]]
*[[Lidstrom:French Press|French Press]]
*[[Lidstrom:Sonicator|Sonication]]
*[[Lidstrom:Sonicator|Sonication]]
 +
*[[Lidstrom:Bug Buster|Bug Buster]]
*[[Lidstrom:Ultracentrifuge|Ultracentrifuge]]
*[[Lidstrom:Ultracentrifuge|Ultracentrifuge]]
Line 86: Line 112:
===General===
===General===
-
*[[Lidstrom: Plate Reader| Plate Reader Setup]]
+
*[[Lidstrom:Enzyme Assay Basics|Enzyme Assay Basics]]
-
*[[Lidstrom: Flow Cytometer| Flow Cytometer]]
+
*[[Lidstrom:Enzyme Assay Data Analysis|Enzyme Assay Data Analysis]]
 +
*[[Lidstrom:Solution Stock Info|Solution Stock Info]]
 +
*[[Lidstrom:Path Length in microwell plates|Path Length in 96-well plates]]
 +
*[[Lidstrom:Reducing Agents|Reducing Agents]]
 +
 
 +
=== Equipment ===
 +
*Plate readers:
 +
**[[Lidstrom: Molecular Devices Plate Reader| Molecular Devices (beige) Plate Reader Setup]]
 +
**[[Lidstrom: Tecan Plate Reader|Tecan (dark grey) Plate Reader]]
 +
* Cuvette spectrophotometer:
 +
** [[Lidstrom:UV-2401PC UV-Vis recording spectrophotometer|UV-2401PC UV-Vis recording spectrophotometer]]
 +
* Centrifuge:
 +
**[[Lidstrom:Sorvall Centrifuge|Sorvall centrifuge]]  goes faster than the regular centrifuges that allow 50mL tubes
 +
* Flow Cytometry:
 +
**[[Lidstrom: Flow Cytometer| Flow Cytometer]]
===in vitro pathway assays (CO<sub>2</sub> project)===
===in vitro pathway assays (CO<sub>2</sub> project)===
Line 93: Line 133:
*[[Lidstrom:in vitro Pathway Assay: Crude Extract Prep| Crude Extract 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)]]
*[[Lidstrom:in vitro Pathway Assay: Sample Analysis via UPLC-MS/MS | Sample Analysis via UPLC-MS/MS (in progress)]]
 +
 +
==== Misc ====
 +
*[[Lidstrom:Assaying enzyme libraries in 96-well plates|Assaying enzyme libraries in 96-well plates]]
 +
*[[Lidstrom:Assaying enzyme libraries|Assaying enzyme libraries]]
== Metabolomics ==
== Metabolomics ==

Current revision

Back home: Lidstrom

Contents

General Lab:

Cell Culture:

Methylotrophs

E. coli

OD measurements

DNA/RNA:

DNA

PCR

Isolation

Manipulation

Fancy Manipulation

Restriction Cloning

Gibson Cloning

Obervation

Mutagenesis

Misc.

Protein

Purification

Analytics

Protein Concentration Determination

Chemicals

Buffers

E. coli

E. coli transformation

Lysing & Cells & Clarifying Extract

Assays

General

Equipment

in vitro pathway assays (CO2 project)

Misc

Metabolomics

CO2 project

Personal tools