The first weeks of the summer fellowship in the VDS stream was a bit hectic. They consisted of learning protocols and techniques. Three plates of PNiC-BSa4 colonies were grown by my partner and me that week. These were later used by the majority of lab students who had reached the cloning stage. My first PCR with plasmid pgbr22 failed, but the second was successful. PCR with pmCherry had to be completed 3 times to get a successful result and PNIC-BSa4 PCR had to be completed twice to get a successful result. The three practice PCRs were completed the 5th week of lab. Protein expression using PfDXR was started in week 3 and the sample of protein was purified the following week. I received my target protein Rickettsia prowazskii FabG …show more content…
The PCR2 sample from the previous week was purified through PCR clean up and nano-dropped. The concentration was 68ng/ul. The concentration of this sample was low (<100ng/ul) because I had skipped the first step in the cleanup process that prepares the column filter for maximum DNA binding. PCR2 and PCR clean up were completed again and the concentration of the sample was averaged to be 147.9ng/ul. Since the concentrations of the samples were adequate, I moved onto PNiC-BSa4 cloning and prepared a sample to be DNA sequenced. RE digest was completed on two PNIcBSa4 prep samples, with the gel indicating that both samples were successfully cut and nano-drop results indicating high purity levels with low concentrations (~32ng/uL). Results from sequencing were nucleotide blasted and confirmed to be PNiCBSa4. I talked to a mentor about my samples and was told that I probably needed a higher concentration ≥100ng/uL, for cloning to be successful. In order to increase the concentration 4 samples of 50uL each were cut and PCR clean up into one sample that had a concentration of 123.5ng/ul. The cloning protocol was completed that week and when the plated were checked, no colonies were present on either of the two plates. My first attempt at cloning failed. The process was completed again, and the sample concentration was averaged to be 112.1ng/ul. The second time, only one
According to my DNA analysis when inputted into the BLAST system my unknown bacteria has an eighty-six percent similarity to Micrococcus Luteus. These result deviate a bit from the results obtained in the Gideon experiment. Since, my DNA sequence result was not as high as I would have preferred this could be due to the limited amount of DNA concentration available after the PCR experiment was completed. A factor that could have allowed me to get a different bacteria species from the biochemical test could have been that the sample taken from the culture could have been contaminated. If the bacteria were not cultured properly this could also affect the results. Making a mistake during the PCR experiment
Freedom Summer was a nonviolent effort by civil rights activists to integrate Mississippi's segregated political system during 1964. It raised the consciousness of millions of people to the troubles of African-Americans and the need for change. Americans all around the country were shocked by the killing of civil rights workers and the brutality they witnessed on their televisions. For nearly a century, segregation had prevented most African-Americans in Mississippi from voting or holding public office. Segregated housing, schools, workplaces, and public accommodations denied black Mississippians access to political or economic power.
Therefore the ratio of A260/230 is the concentration of DNA/ the concentration of other containments (Wilfinger). From these values one can determine that the sequencing process was successful. These values displayed that the sequencing reaction worked, but did not give the desired results and did not have a concentrated product. The amount of the DNA sample and water had to be adjusted in the tube that was going to be sent to the Cornell lab. The amount that was supposed to be used was 1.5ul of mtDNA. This value was determined by 100ng of DNA required x volume of DNA in ul divided by the concentration of DNA. The instructor simply advised to double the amount of mtDNA that would be placed in the tube. The volume of DNA used was 3uL, and the volume of sterile water used was 13.4uL. The volume of the primer used was 1.6uL(which was not adjusted). Figure 2 displays the reverse complement of the mtDNA that was obtained from the Cornell lab sequencing the mtDNA sent to them. This was then used to compare it with the CRS mtDNA. Figure 3 displays the aliment of the CRS and the revers compliemt of the mtDNA sent to the Cornell
In order to talk about the Freedom Summer project, we first have to identify it’s roots and the history behind it. Before the Freedom Summer project, there was the civil right movement in which thousands of African Americans protested for equality. Equality didn’t mean the term referred in the court case Plessy vs Ferguson “separate but equal.” African Americans wanted to end the era of segregation, this include not having to use lower quality public facilities than whites, not having to give their seats in a bus if a white person wanted to sit there, and the right to vote. With the help of different civil right activist such as Bayard Rustin, Jo Ann Robinson, Martin Luther King, Rosa Parks, and all the people that marched and protested against
In the Summer of 1964, the Mississippi Summer Project, also known as Freedom Summer, was organized by several Civil Rights organizations such as the Student Nonviolent Coordination Committee and the Congress on Racial Equality. The event that caused the start of Freedom Summer were that many African-Americans were not registered to vote, this was because the southern states had implemented literacy tests that were unfair and could be interpreted differently, and Poll Taxes which were ridiculous amounts to pay ("Freedom Summer"). Other causes include the case of Plessy Vs. Ferguson, which stated the “Separate but Equal” clause, and also Racial segregation. Freedom Summer Volunteers included White Northerners, and the organization was made to focus on the Mississippi Freedom Democratic Party, Voter Registration, Freedom Schools, and Community Centers (“Timeline: Freedom Summer”). There was a need for the Freedom Summer because Many African-American People in the south were being oppressed by Literacy Tests and Poll Taxes.
June 21, 1964, the Freedom of Summer! Civil rights organization included the Congress on Racial Equality and Student Nonviolent Coordinating Committee, which a voter signing up to drive, or Freedom Summer, Aimed to increase voters in Mississippi. Freedom Summer has most of black Mississippians and over one-thousand out of state white volunteers face abuse and harassment from Mississippi's white population. The Ku Klux Klan police did a series of violent attacks towards them. About a hundred white college students had help cofo register voters on November 1963, and several hundred students more had been invited to expand the voters project in the Freedom Summer.
Although the experiment was as fair as it could have been, there were some factors that were beyond our control. Firstly, the tubers that we used may not have been from the same specimen,
The objective of this experiment was to identify two unknown bacteria from a mixed culture. Which was done by using the aseptic technique which was very important to avoid any contamination and keeping the workspace clean while culturing bacteria for different tests. To start, I chose a tube which had a solution of mixed culture. I used the flame to sterilize the inoculating loop and dipped it into the tube and streaked for isolation on 2 TSA plates and placed them in an incubator at 37 for 24 hours. Next day I observed the growth of 2 different types of colonies one for each unknown on the two plates. So I picked the best one and labeled it as master plate and discarded the other plate. From the master plate, I subcultured each type of colony
Summer school is beneficial to students. it Promotes social development and personal growth. It encourages physical activity and routine. Students get in low income groups when they don't go to summer school because they forget stuff. Summer school is beneficial to students.
Weekly Research- Week 2- A & P III -Tamara Goins- Antonelli College- Ericca Peacock- Instructor
Even though we had followed procedure and accurately mixed the correct amount and type of DNA and Master Mix together, as shown in Table
Our results had been inconclusive in this part of the lab. The unknown was run with the other class due to the loss of a PCR tube. The tube had been redone and run in the gel and results had been inconclusive. No results had been shown for our gel. Meaning something along the way had been done incorrectly and as a result the experiment had no results. It is possible that primers or something was not added to the tube causing failed results. A wide variety of things could have gone wrong. But what we do know is that results should have occurred. In figure #6 the gel that was done by our class we could see that there are results. There is a very noticeable band that shows that this experiment could be done and that it wasn’t any machines fault but human error. The class had a band around 1500 base pairs which would be completely correct because 16s rRNA gene is usually 1500 base pairs long (Clarridge
While trying to make MPI I ran out of my MTL. I had to go back to phage titer procedure in order to flood it and get MTL. I started off with normal phage titer procedure. I got an MTL. From that MTL I did several MPIs and those didn’t yield proper results. I ran out of my MTL and had to go back to the phage titer procedure. At this point, my plaques were not showing. So, I made little changes with my procedure where I picked up several plaques and using the original amount of PB (50µl). However, plaques still weren't showing up, so I decreased the amount of PB to 30µl. & continued to pick several plaques (instead of one plaque) in order to get a more concentrated 100 tube. It Resulted in some small plaques, but nothing even close to a web pattern.
Starting May 22, 2017 I will be attending a summer class for school from 6PM-9:45PM that requires me to be absent from Monday night drills and meetings. The class is from Monday-Thursday for six weeks. With this being said I would like permission to be excused from drills, meetings, etc. until June 30, 2017.
Experiment 4 & 5: PCR did not work for my obtained DNA as there was no evidence of movement of the DNA. PCR did work for lab numbers 33 and 12. The size of my fragment could not be determined and cannot compare to the size that was expected. The negative control was clean as there was no traces of movements.