Experiments

Overview

You will find below all the protocols we conducted during the year. For convenience, we organized them in three sections:

• Plasmid construction

• Protein isolation and characterization

• Solution testing

Plasmid construction

Competent bacteria

Competent bacteria transformation

DNA miniprep using Monarch kit from NEB

Double Digestion with restriction enzymes

Electrophoresis (Gel preparation and migration)

Gel Extraction by using Monarch DNA gel extraction kit from NEB

Ligation protocol using NEB reagents

Midi prep with NucleoBond Xtra Midi kit of Macherey-Nagel

Polymerase Chain Reaction using Q5 High Fidelity 2X Master Mix of NEB

Preparation of Petri dishes

Golden Gate Assembly

Protein isolation and characterization

Bicinchoninic Acid Assay (BCA)

Total protein extraction

Electrophoresis of proteins (SDS-Page)

TCA protein precipitation

TMV Coat protein collection and purification

TMV Mechanical Inoculation

TMV Virion purification

Dialysis of TMV Coat Protein

RNA encapsidation in TMV Coat Proteins

Solution testing

Brilliant III Ultra-Fast qRT-PCR Master Mix

Total RNA extraction from Plants

Purification of siRNA

Specific siRNA production detection

Purification of dsRNA