User:Elizabeth Ghias/Notebook/Experimental Biological Chemistry/2011/09/21: Difference between revisions
From OpenWetWare
No edit summary |
No edit summary |
||
Line 25: | Line 25: | ||
# The DNA samples were prepared for the gel by mixing 1 μL of loading dye with 5μL of PCR product. | # The DNA samples were prepared for the gel by mixing 1 μL of loading dye with 5μL of PCR product. | ||
# 6 μL of each sample was added to each gel wall, beginning with a DNA ladder in the first well on the left. | # 6 μL of each sample was added to each gel wall, beginning with a DNA ladder in the first well on the left. | ||
<u>Protein Expression</u> | |||
# The starter culture was centrifuged at 4500 rpm for 15 minutes. | |||
# The cell pellets were resuspended in 4 mL of Luria Broth. | |||
# The resuspended cells were divided between the four Fernbach flasks to inoculate the expression culture media. | |||
# The expression cultures were incubated at 37°C until their absorbance at 600 nm equaled 0.6. | |||
# 1 mL of the IPTG prepared the previous day was added to each flask. | |||
# The flasks were shaken for 2 hours. | |||
# Each flask was spit into 2 750 mL plastic containers with caps, so that each plastic container had the same mass of expression culture. | |||
# Four of the plastic containers were centrifuged for 15 minutes each at 4500 rpm. | |||
# | |||
Revision as of 14:46, 27 September 2011
Experimental Biological Chemistry | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
ObjectiveTo preform electrophoresis using an agarose gel to determine if the PCR performed the previous day successfully mutated the GFP protein. To carry out protein expression using the starter culture media and the expression culture media prepared the day before. DescriptionElectrophoresis Procedure
Data
NotesThis area is for any observations or conclusions that you would like to note.
|