What size fragments would you expect when cutting the plasmid with the two enzymes used
DO NOT DO RESULT SECTION
This lab report will cover the transformation of E.coli and the restriction digest of the plasmid (labs 5 to 8).
Use the Standard lab report format and include the following
– Overall objective
– Background to what you were trying to do
– Methods – used to create reagents and used for the transformation, plasmid isolation, restriction digest and agarose gels. Create sub section for each
Transformation
– What was the selectable marker?
– How do you know you successfully transformed the cells with pGlo
– What was the most critical step in the transformation process
Plasmid preparation
– Why does only the E. coli grown in ara/amp fluoresce
Restriction digest
– What size fragments would you expect when cutting the plasmid with the two enzymes used?
– Are the results of the plasmid what you expected.
– Are the two fragments the sizes you expected ?
This lab report will cover the transformation of E.coli and the restriction digest of the plasmid (labs 5 to 8).
Use the Standard lab report format and include the following
– Overall objective
– Background to what you were trying to do
– Methods – used to create reagents and used for the transformation, plasmid isolation, restriction digest and agarose gels. Create sub section for each
Transformation
– What was the selectable marker?
– How do you know you successfully transformed the cells with pGlo
– What was the most critical step in the transformation process
Plasmid preparation
– Why does only the E. coli grown in ara/amp fluoresce
Restriction digest
– What size fragments would you expect when cutting the plasmid with the two enzymes used?
– Are the results of the plasmid what you expected.
– Are the two fragments the sizes you expected ?
| Biol 390-Lab 5 Preparation for Transformation |
2
| Objective | · Prepare materials for transformation · Prepare TAE buffer for electrophoresis in a later lab. |
| Background | Transformation of bacterial with plasmids is a fundamental technique in genetic engineering. Cells can be made competent to take up plasmids by treatment with divalent cations and heat. Transformed cells will contain a plasmid that contains a gene that confers resistance to the antibiotic, ampicillin. Therefore, transformed cells can be selected by growth on LB/amp plates. Growth on LB/ara /amp will cause the product to be expressed as the plasmid contains an arabinose promotor |
