Red Florescent Protein Lab
Purpose
To make RFP from jelly fish in bacteria and learn about the steps of this process and genetic engineering.
To make RFP from jelly fish in bacteria and learn about the steps of this process and genetic engineering.
Materials and Procedure
2A - Materials and procedure in Amgen lab manual part 2A
4A - Materials and procedure in Amgen lab manual part 4A
5A - Materials and procedure in Amgen lab manual part 5A
6 - Materials and procedure in Amgen lab manual part 6A
2A - Materials and procedure in Amgen lab manual part 2A
4A - Materials and procedure in Amgen lab manual part 4A
5A - Materials and procedure in Amgen lab manual part 5A
6 - Materials and procedure in Amgen lab manual part 6A
Experimental Overview
Part 2A: Verification of plasmid by restriction digest. We cut out the RFP-ara from the bacterial plasmid with BanH 1 and Hind III in order to get the RFP so that we could make sure we had all the components we needed.
Part 4A: Verification of plasmid by electrophoreis. We just analyzed what we did in 2A in order to see if we did everything right.
Part 5A: Transformation of bacteria with recombinant plasmid. We put the protein we extracted in part 2A into bacteria so that it could be produced.
Part 6: Purification of RFP using chromatography. We just purified the RFP from the bacteria.
Part 2A: Verification of plasmid by restriction digest. We cut out the RFP-ara from the bacterial plasmid with BanH 1 and Hind III in order to get the RFP so that we could make sure we had all the components we needed.
Part 4A: Verification of plasmid by electrophoreis. We just analyzed what we did in 2A in order to see if we did everything right.
Part 5A: Transformation of bacteria with recombinant plasmid. We put the protein we extracted in part 2A into bacteria so that it could be produced.
Part 6: Purification of RFP using chromatography. We just purified the RFP from the bacteria.
Results
Part 2A: We were able to to verify that all the components were there. Part 4A: See Left Part 5A: The protein reproduced well enough for our purposes. Part 6: We got our RFP to show up at the end. Data Analysis
Everything seems to have gone well and we got our red fluorescent protein to show up. We had all the components we needed, the protein reproduced quite a bit and all the reproduced proteins showed the red coloring. |
Gel Analysis
Our class' gels failed to show the RFP, so we borrowed gels from another class and we're going to analyze them. The lane I'm analyzing is the last one to the right. This lane is pretty pure, but not completely because there's traces of other proteins in the gel. This is most likely due to when they were doing the chromatography they either didn't wash the tube completely or they accidentally collected some of the washing fluid with their RFP. The RFP is supposed to be 28 kD but their's is only about 17 kD. |
Reflection
This lab went really smoothly, at least compared to the other labs. I feel like I understand what was going on when it did, and my pipetting skills are pretty good now. I did not have any complaints about the lab except for just getting lost in what we were supposed to do, but that's our fault. I found this lab to be easier, maybe, and I was interested in what was going on most of the time.
This lab went really smoothly, at least compared to the other labs. I feel like I understand what was going on when it did, and my pipetting skills are pretty good now. I did not have any complaints about the lab except for just getting lost in what we were supposed to do, but that's our fault. I found this lab to be easier, maybe, and I was interested in what was going on most of the time.