Saul+H.+(springer?)

=__**Week 14,15**__= Three trials for Clean up for PCR squared Eluted in Tris HCl in order to get rid of enzymes, dNTPS and any unnecessary contamination, to purify the sample. move on to cloning, inserting gene into vector, and further advance in my research.
 * Analysis**: concentrations of these three trials is enough, all around 170ng/uL. Able to prepare and cut pNIC-Bsa4 and

= = =__Week 11,12,13__=
 * 12042014- Need more analysis of work completed in the lab**

1162014- Good Job =__Week 8,9,10__=
 * Analysis**: able to get my primary and secondary on working this confirms elongation of my gene and over replication of it. Will now be moving up to my PCR squared for purification and over amplification.

9232014- Good job =**__Week 5,6&7__**=
 * __10/3/14 Primary PCR Trial 2__**
 * __10/2/14 Primary PCR trial1__**

Purpose of primary PCR is to put together my oligo mix which partially makes a double strand but gene is not fully assembled. Gene development is verified after running a gel of my pcr mix with a smear shown, but so far i have not been able to get a smear on my gel =** __9/30/14 Plasmid Midi-Prep__ **= Midi Prep was done for extracting/purifying pNIC BSa4 out of pellet, and remove unwanted cell components.
 * __Analysis:__**
 * Analysis:**

__**Analysis**__: pNIC-Bsa4 plasmid was virtually cut with Bsal using NEB cutter. Image of this cut was made and put above as well. Gel of this cut was made too with 1 kb ladder and the gene segments of bsal on the plasmid. The sequence that Bsal cut was removed and my CDS of pF6Fg2 sequence was inserted for cloning.
 * __9/29/14 PCR Primer Design Tails for pNIC-Bsa4 Cloning__**

=**__Week 3&4__**=
 * 9/15/14 PCR**


 * Analysis:** the purpose here for this lab was to amplify purple protein coding sequence in the pGBR22 plasmid using forward and reversed primers. dilutions were made for the tubes. Tube A had a 1:10000, tube B had a 1:1000, tube C had a 1:100. After PCR, gels were made to analyze the samples to see the DNA bands. The gel was run for about 40 mins, at 105 volts until the visible blue marker dye was near the bottom 1/4th of the gel. After gel was run nothing appeared. Unfortunately the gel only showed the two DNA ladders that were placed on lanes 2 and 7. These errors were systematic errors because most of the gel running machines do not work so this being are probable cause of why the gel did not run appropriately. Next possible steps would be to see if one of my lab peers managed to run a gel and get reasonable date out of them so that i can see it and get comparable conclusion for my samples.


 * 9/9/14 RE Digest Day 1**






 * Analysis**: the purpose for this lab was to make plasmid digestion using EcoRi and PVUII and a combination of both. After the tree samples were prepared, we ran a gel with 1 kbDNA ladder, our three samples and uncut plasmid in a total of five lanes. The gel was run at 100V, until blue dye was half way since DNA id negatively charged for about 40 min. The Bands did appear as i expected even though the lanes were kind of blurry and couldn't tell whether they had other components in them. Next steps would be to analyze the gel with people of greater experience to confirm my gels validity.


 * Day 2 Transformation 9/8/14**


 * Day 3 Transformation 9/9/14**



For day 2 and days 3 of transformation bacterial colonies with plasmid pNICBsa4 were overgrown and stored for later usage when we begin Mid Prep. The purpose for day 2 was to grab a bacterial colony and overgrow it over night for about 16 hrs in 37 shaker at fast rpm. Day 3 the contents of the flasks were put into conical tubes were they were spined down in 4 centrifuge at 6000xg for about 15 mins.No contamination in pallets.
 * Analysis**:


 * 982014- Nice work, remember to have all your pictures next** **time**

= = =__**Week 1&2**__=



**__Analysis__**: Our transformation of DH5alpha E.Coli did happen and it grew as you can see on the figures above. Whether the plasmid was inserted into the bacteria’s DNA can not be know but there were definitely colonies of bacterial in both agar plates. The next step would be to continue with the lab and over grow the bacteria and see whether the bacteria can expressed our desired protein.



__**Analysis**__: The purpose for this lab was to verify DNA purity using nanodrop. Unfortunately we forgot to gather the second graph and were only able to salvage the first one. The recorded concentration was 302.9 ng/ul for the pgbr22 and for the ratios 260/230 and 280/230 the readings were 1.88 and 2.34 respectively, which are close enough to pure pgbr22 which have 1.8 and 2.1 for the ratios 260/230 and 280/230. This validates the sample of pgbr22 and we are able to proced to the DNA sequnceing facility for them to sequence our sample for further use.