Team:Warsaw/Project
From 2011.igem.org
Project description
This year's goal of iGEM Warsaw team is to develop 2 foundational
techniques for synthetic biologists:
Synthetic Cloning
Our first goal is to make easy and efficient protocol for cell free
cloning. It skips most basic step of classical cloning approach -
plasmid propagation in bacteria or yeast. This allows cloning of
constructs that are toxic to host cells (i.e. nucleases or lysins) and
speeds up the whole procedure at least three times. The cell free
cloning process involves rolling circle amplification of ligation
products by phi29 polymerase. Our protocol is speed-oriented so we'll
try to optimize all time consuming steps: DNA digestion, substrate
purification, ligation and product amplification. We'll also try to
deal with specificity problems of phiX29 polymerase by designing RNA
primers optimized for BioBrick amplification.
Expression Adaptors
Additionally we are working on expression adapters that would provide standardized protein expression from each RBS from community collection. Also we are working on adapters that increase protein expression