McClean:Protocols: Difference between revisions
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==Health and Safety== | ==Health and Safety== | ||
*[[McClean: Safety101 | Safety 101]] | *[[McClean: Safety101 | Safety 101]] | ||
*[[McClean: SharpsDisposal | Sharps Disposal]] | |||
*[[McClean: OkToTrash | Ok to Trash Stickers]] | *[[McClean: OkToTrash | Ok to Trash Stickers]] | ||
| Line 88: | Line 89: | ||
*[[McClean: Anneal and Extend | Anneal and Extend]] | *[[McClean: Anneal and Extend | Anneal and Extend]] | ||
*[[McClean: Designing "Yeast Toolkit" compatible primers | Designing "Yeast Toolkit" compatible primers]] | *[[McClean: Designing "Yeast Toolkit" compatible primers | Designing "Yeast Toolkit" compatible primers]] | ||
*[[McClean: Q5 Mutagenesis Kit from NEB | Q5 Mutagenesis Kit from NEB]] | |||
== | ==Modular Cloning-Yeast Toolkit assembly== | ||
*[[McClean: Designing "Yeast Toolkit" compatible primers | Designing "Yeast Toolkit" compatible primers]] | |||
*[[McClean: Golden Gate-Making a new part | Golden Gate-Making a new part]] | *[[McClean: Golden Gate-Making a new part | Golden Gate-Making a new part]] | ||
*[[McClean: Golden Gate- Making a multi-part plasmid | Golden Gate-Making a multi-part plasmid]] | *[[McClean: Golden Gate- Making a multi-part plasmid | Golden Gate-Making a multi-part plasmid]] | ||
| Line 135: | Line 138: | ||
*[[McClean: Phosphate Buffered Saline| Phosphate Buffered Saline (PBS, 10x)]] | *[[McClean: Phosphate Buffered Saline| Phosphate Buffered Saline (PBS, 10x)]] | ||
*[[McClean: Potassium Phosphate | Potassium Phosphate Buffer]] | *[[McClean: Potassium Phosphate | Potassium Phosphate Buffer]] | ||
*[[McClean: Plasmid Prep Buffers | Generic recipes for Qiagen buffers]] | |||
*[[McClean: SeventyPercent_EtOH | Seventy Percent (70%) Ethanol]] | *[[McClean: SeventyPercent_EtOH | Seventy Percent (70%) Ethanol]] | ||
*[[McClean: TAE_50X | TAE Buffer (50X)]] | *[[McClean: TAE_50X | TAE Buffer (50X)]] | ||
| Line 164: | Line 168: | ||
==Equipment Maintenance== | ==Equipment Maintenance== | ||
*[[McClean: RollerDrum_BrushReplacement | Roller Drum Brush Replacement]] | *[[McClean: RollerDrum_BrushReplacement | Roller Drum Brush Replacement]] | ||
*[[McClean: PlasmaPreen_ORingReplacement | Plasma Preen O-Ring Replacement]] | |||
==Misc== | ==Misc== | ||
Revision as of 10:15, 23 May 2017
Back to McClean Lab
Basics
- Lab Expectations
- Lab Rules
- Lab Jobs
- Joining the Lab
- Leaving the Lab
- Lab Notebooks
- Dilution of Oligos
- Ordering Media
- Ordering Supplies
- Protocol Template
- Lab Database
- Lab Organization Notes
Introductory Exercises
Health and Safety
General Lab Procedures
- Washing Glassware
- Dry Ice-Ethanol Bath
- Pouring Gels for Electrophoresis (Mike)
- Using the Nanodrop2000
- Quantifying DNA with the Qubit Fluorometer
- Ordering Liquid N2 for Common Lab
Yeast
- Drug Concentrations
- Basic Chemostat Guide
- Bayanus Transformation
- Cleaning Floating Pin Replicators
- Fluorescence in situ Hybridization (Colin)
- Fluorescent in situ Hybridization (Ping)
- Fluorescent in situ Hybridization (Gasch Lab Collaboration)
- Fixation of Yeast (Bisaria Protocol)
- Fixation of Yeast (McClean Protocol)
- Fixation of Yeast (P. Xu Protocol)
- Frogging a Serial Dilution
- Frogging Tetrads
- Genomic DNA Prep (Bust 'n' Grab Protocol)
- GEV Strain Construction
- Glycerol stocks (yeast)
- Hemacytometer protocol for yeast
- Making and Using Frozen Yeast Competant Cells
- Mating Type Testers
- Phluorin Calibration
- Pinning (96 or 384 format)
- Plasmid Loss Assay
- Random Spore Prep
- Sanger Sequencing
- Smartstat Startup
- Sonication of Yeast
- Sporulation
- Tetrad Dissection
- URA Pop-out
- Yeast Dubious ORFs
- Yeast Mating Halo Assay
- Yeast Nomenclature
- Yeast Recombinational Cloning
- Yeast Transformation (S. cerevisiae)
- YMC Induction in Chemostat
- Zygote Picking
- Working with the LoxP/Cre System in S. cerevisiae
- Western Blot
- Membrane Stripping
- Membrane Stripping-Mild
- Invasive growth assay
Bacteria
- Transformation of E. Coli
- E. coli Glycerol Stocks
- E. coli Competent Cells
- Colony PCR (E. coli)
- E. coli Electroporation
- Plasmid Prep from E. coli using a Qiagen Kit
PCR
- Designing Primers (Yeast)
- Colony PCR (Yeast)
- dNTP Stocks
- Touchdown PCR
- Takara PrimeStar PCR
- Anneal and Extend
- Designing "Yeast Toolkit" compatible primers
- Q5 Mutagenesis Kit from NEB
Modular Cloning-Yeast Toolkit assembly
- Designing "Yeast Toolkit" compatible primers
- Golden Gate-Making a new part
- Golden Gate-Making a multi-part plasmid
Flow Cytometry
- General Flow Cytometry Procedure
- Cycloheximide Concentration Assay
- Flourecent Protein Folding Time Assay
- Induction time course
- Phase Plane Mapping
Media
- KS Amino Acid Supplement
- LB1
- LB2
- Low Fluorescence Media
- Low Fluorescence Agar Membranes
- Magic Marker Medias
- Quickie YPD supplemented with Adenine
- Synthetic Complete (SC) Yeast Media
- Synthetic Complete (SC) Media w/Drugs
- Synthetic Complete (SC) Media w/Canavanine
- YNB1
- YNB2
- YNB3
- YNB4
- YPD2
- YPD4
- Chloramphenicol media
- Kanamycin media
- 5FOA
Stock Solutions
- Agarose for gels
- α-Factor 1mg/ml Stock
- Bacterial Glycerol 65%
- Carbenicillin Stock Solution
- Concanavalin A Solution without ions
- Cycloheximide Stock Solution
- DNA Ladder Stock for PCR
- EDTA (0.5M)
- Geneticin/G418 Stock Solution
- Glucose Solution for Media
- PBS/0.1%Tween
- Phosphate Buffered Saline (PBS, 10x)
- Potassium Phosphate Buffer
- Generic recipes for Qiagen buffers
- Seventy Percent (70%) Ethanol
- TAE Buffer (50X)
- TE Buffer (10X)
- Tris Buffer
- Yeast Glycerol 30%
- Chloramphenicol
- Kanamycin
Microfluidics
- Concanavalin A Solution for Microfluidics
- Constructing PDMS Flow Cells
- FCS2 chamber for Metabolic Cycling
- Bandwidth Experiments
- Bioptechs Temperature Experiments
Microscopy
- Introduction to the microscope
- Microscope Dos and Don'ts
- Nikon TI Basic Use
- Registering objectives on the Nikon
- 96 Well Plate Assay
- NIS Elements Repair
- New Scope Settings
- Quickie ImageJ Quantification
- NikonTI-Eclipse Setup and operation using Micromanager
- Cleaning the Camera's Cover
Equipment Maintenance
Misc
- 96 Well Plate Print Out
- AddGene Orders
- Blue Light Overview
- Sequencing
- Bibtex4Word (Add-in for word that allows you to use your existing BibTeX database)
- Orders from IDT/Genewiz/Macrogen/GenScript
- Searching a Sequence for All Database Primers
- Annealing Oligos
- Oligonucleotide phosphorylation, Annealing and Ligation
- How to Connect Blue LED to a Power Source, Parts and Instructions
- Compiling Kappa in a Linux environment
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