Team:UAM Poznan/Experiments

UAM_Poznan Team

METHODS

MEDIA PREPARATION AND BACTERIAL CULTURES

LB medium (Lysogeny-Broth)

2xLB medium (2xLysogeny-Broth)

SB-PKB medium (Super Broth-PKB)

SOC medium

5xM9 salts

M9 medium

Plates with M9 medium

Preparation of electro-competent Escherichia coli cells

Transformation of Escherichia coli via electroporation

E.coli glycerol stocks


CONSTRUCT DESIGN AND CLONING METHODS


DNA amplification

Electrophoresis of DNA fragments in agarose gels

Gel extraction

Chromosomal DNA isolation from Escherichia coli

Plasmid isolation and purification

Construct verification by colony PCR and Sanger DNA sequencing


CONSTRUCT AND PART DESIGN


Primers design

Construct assembly by CPEC

Coding sequence optimization

ORFs

Codon context optimization


MEASUREMENTS

Fluorescence measurements

OD600 Measurements



References
1. Koressaar T1, Remm M. (2007) Enhancements and modifications of primer design program Primer3. Bioinformatics 23(10):1289-91.
2. Untergasser A1, Cutcutache I, Koressaar T, Ye J, Faircloth BC, Remm M, Rozen SG. (2012) Primer3—new capabilities and interfaces. Nucleic Acids Res. 40(15):e115.

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