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From 2014.igem.org

Contents 1 Protocols 1.1 Media, Plates and Solutions 1.1.1 CCMB 1.1.2 LB-Agar 1.1.3 Luria Broth 1.1.4 Super Optimal Broth (S.O.B.) 1.1.5 Guanidinium Hydrogen Chloride 1.1.6 YPD 1.1.7 Yeast Liquid Cultures 1.1.8 PBSF 1.2 Basic Cloning 1.2.1 Polymerase Chain Reactions 1.2.2 Restriction Endonuclease Reaction (Digestion) 1.2.3 Ligation 1.3 Escherichia coli Protocols (XL1-Blue and XL10-Gold) 1.3.1 Competent Cell Culturing 1.3.2 Competent Cell Transformations 1.3.3 Plating 1.3.4 Overnights 1.3.5 DNA-Extraction and mini-preps 1.3.6 Glycerol Stocks 1.4 Saccharomyces cerevisiae (PYE1 Yeast) 1.4.1 Transformations 1.4.2 OVernight Culturing and Passaging 1.4.3 Glycerol Stocks 1.5 Flow Cytometry and Fluorescence Activated Cell Sorting 1.5.1 Dilutions 1.5.2 Final Preparations 1.6 Protein Expression 1.6.1 Overnight Cultures 1.6.2 Protein Extraction and Purification 1.6.3 Nickel Nitrotriacetic Acid Chromatography 1.6.4 Size Exclusion Chromatography (S.E.C.) 1.7 Stability Analysis 1.7.1 Thermal Melts 1.7.2 Guanidinium Hydrogen Chloride Melts

Protocols

Media, Plates and Solutions

CCMB

LB-Agar

Luria Broth

Super Optimal Broth (S.O.B.)

Guanidinium Hydrogen Chloride

YPD

Yeast Liquid Cultures

PBSF

Basic Cloning

Polymerase Chain Reactions

Restriction Endonuclease Reaction (Digestion)

Ligation

Escherichia coli Protocols (XL1-Blue and XL10-Gold)

Competent Cell Culturing

Competent Cell Transformations

Plating

Overnights

DNA-Extraction and mini-preps

Glycerol Stocks

Saccharomyces cerevisiae (PYE1 Yeast)

Transformations

OVernight Culturing and Passaging

Glycerol Stocks

Flow Cytometry and Fluorescence Activated Cell Sorting

Dilutions

Final Preparations

Protein Expression

Overnight Cultures

Protein Extraction and Purification

Nickel Nitrotriacetic Acid Chromatography

Size Exclusion Chromatography (S.E.C.)

Stability Analysis

Thermal Melts

Guanidinium Hydrogen Chloride Melts

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