"
Team:GeorgiaTech/Project/Primers
From 2014.igem.org
Biobrick Primers for Site-Directed Insertion Mutagenesis
Ribosomal Binding Site Insertion
Purpose
The RBS primer is a standard primer that can be used for site directed mutagenesis in all linearized backbones available from the iGEM database.
Background
Most of the BioBrick parts requires a Ribosomal Binding Site to express the protein. This has proven problematic as the RBS is very small and hard to insert using 3A Assembly. We believe the creation of an oligo primer with this RBS may be easier to place the RBS in front of the protein that is needed to be expressed. An easy oligo primer that could be used in PCR extension and then placed into a vector backbone would save time and frustration as to whether the part had been inserted.
Promoter Sequence Insertion
Purpose
The Promoter primer is a standard primer that can be used for site directed mutagenesis in all linearized backbones available from the iGEM database and the Anderson RBS sites from the Anderson RBS library.
Background
Most of the BioBrick parts requires a Promoter to express the protein. This has proven problematic as the promoter is very small and hard to insert using 3A Assembly. We believe the creation of an oligo primer with this promoter may be easier to place the promoter in front of the RBS site that is needed to be expressed. An easy oligo primer that could be used in PCR extension and then placed into a vector backbone would save time and frustration as to whether the part had been inserted.
