Plasmid Lab Report Worksheet ( 50 pts)
Section 002 Date November 19th 2011
Comparing Plasmids pBR 322 and pGFP
Introduction Define genetic engineering and cloning. Describe the two most important tools needed for cloning and genetic engineering techniques, and how the two tools will be used in this lab. 3 pts
Genetic Engineering is the technique and process of manipulating an organisms genome to alter its phenotype or characteristics, and producing genetically identical copies of an organism is called cloning.
Two important tools in genetic engineering and cloning are transformation and electrophoresis. Transformation is a processes and technique of inserting a foreign DNA into an organism’s genome. The second important tool, electrophoresis, allows us to differentiate DNA sequences through restriction enzyme and DNA fragment bands. In this lab we will transform E. coli bacteria via heat shock and calcium chloride, and use electrophoresis to examine the purified plasmid DNA (Freeman 391.)
Materials and Methods_(1 pt)_____________________________________________
The transformation technique is described in part 1 on pages 87 and 88 in the Lab Manual.
The electrophoresis technique is described in part 3 on page 93 in the Lab Manual.
The plasmid isolation “mini-prep” technique and restriction digests are described on pages 90-92 in the lab manual.
To determine which plasmid is A and which is B using the transformation techniques and analyzing the restriction digests of both re-isolated plasmids.
To understand how genes from different organisms can be recombined, identified, and expressed using plasmids.
To learn the techniques of transformation, plasmid isolation, and gel electrophoresis.
Part I Transformation with pBR 322 and pGFPuv
pBR 322 is a circular plasmid...