Giovanni+O.+(Springer)

=__Week 14 & 15__=

Preparation of pNIC-Bsa4 as Accepting Vector
Analysis: Concentration at 260nm wavelength had an absorbance of 111.5 ng/uL =120042014- Good work, but don't forget a conclusion next time= =__Week 11, 12, and 13__=

PCR cleanup
Analysis: Maximum absorbance was at 260nm which confirms that sample contains DNA. The concentration at maximum absorbance was of 130.3 ng/uL, which is a good concentration. The next step is to move on to cloning.

PCR squared
Analysis: The PCR squared protocol followed was the same as secondary PCR (NEB guidelines) except for the increase from 20 cycles to 30 cycles. The bands are visible in the gel at the right length. Since PCR squared was successful and there didn't seem to be any major errors, the next step was to move on to PCR clean up.

1162014- Where is week 9,10? good job with week 7&8 9232014- You should elaborate on your analysis =__Week 9 and 10__=

Secondary PCR
Analysis: There is a visible band seen were the secondary PCR solution was run. The band is light, however this could have occurred because the ethidium bromide was not mixed correctly, which could have also caused to white spots at the top. The next step is to run PCR squared which will also confirm that secondary PCR worked.

=__Week 7 and 8__=

Secondary PCR 1st and 2nd Trial Re-run
Analysis: Lane five has a barely visible smudge, however, since it is not clear enough, the secondary PCR will be run again. This gel demonstrated that the error was not in the gel, and was in the conditions in which the secondary PCR was run. Therefore the times at which the PCR was run will be changed in the next run to the NEB recommended guidelines.

Secondary PCR 2nd Trial
Analysis: Second trial for secondary PCR was run under the same conditions as the first trial and gave the same results of showing no bands or even a smudge. Errors could have occurred from making the mixture incorrectly or the process of running the gel could have also caused error. To test if the gel was not run incorrectly the first and second trial of secondary PCR will be run again. =__Week 5 and 6__=

Secondary PCR

 * Analysis:** There was clear band shown in the gel from secondary PCR, therefore PCR did give the expected results. Secondary PCR will be ran one more time in the same conditions.

Primary PCR

 * Analysis:** The first gel of primary PCR showed that the PCR procedure did not work. However, when the second gel was ran, it showed that primary PCR had worked on the first trial and not on the second trial.

Plasmid Midi-Prep

 * Analysis:** The average concentration for the two nano drop trails the average concentration was 59.3 ng/ul at the 260 wavelength. This concentration is in the range for what the range should be for pNIC, 30-60.

09232014- Keep up the good work, don't forget to include your captions =**__Week 3 and 4__**=

**PCR**





 * Analysis:** This PCR did not turn out properly. As it can be seen from the gel only the ladder came out correctly. The last lane contains DNA which should have been the one without DNA since it is the control. Lane 2, 3, and 4 don't contain any DNA which should have DNA.

**Restriction Enzyme Digest**

 * Analysis:** The bands did not appear as expected. The ladder did appear correctly and the uncut plasmid lane also appeared. However, the lanes that contains the restriction enzymes did not contain plasmid.

**Day two and three of** **Bacterial Transformation** **of pNIC-Bsa4 //E. coli// DH-5 alpha cells**



 * Analysis:** The pellets showed that the transformation was successful. The plasmid was taken by the bacteria and the pellet was able to be separated from the liquid.

=__**Week 1 and 2**__=

**Bacterial Transformation** **of pNIC-Bsa4 //E. coli// DH-5 alpha cells**




From the results above it can be seen that the bacteria did grow in one day of incubation. It can be observed that more bacteria cultures grew in the agar plate containing 50ul than the agar plate with 10ul of bacteria. The next step would be to grow the bacteria and then isolate the desired protein created by the bacteria.
 * Analysis:**

Nanodrop
Figure 1. First trial of nanodrop absorbance measurement of pGBR22 plasmid



Figure 2. Second trial of nanodrop absorbance measurement of pGBR22 plasmid

The results demonstrated that the maximum absorbance was at the 260nm wavelength. This indicates that the nanodrop solution does in fact contain the DNA, since most DNA absorbs at the 260nm wavelength. After determining the average concentration, which was 241.3 ng/ul, the solution can be prepared and sent to DNA sequencing to confirm that the plasmid is pGBR22.
 * Analysis:**