Embedded Files
5/28/21
5/28/21
Received pKK-mRuby3-TEV and placed the bacterial stab in Fridge B
6/01/21
6/01/21
pKK-mRuby3-TEV was streaked onto two plates (pKK-mRuby3-TEV-1 and pKK-mRuby3-TEV-2). The plates were LB+AMP made the same day. pKK-mRuby3-TEV-1 was performed by LC and the sterile loop was not dipped in ethanol between each quadrant streak. pKK-mRuby3-TEV-2 was performed by HS and the sterile loop was dipped in ethanol between each quadrant streak. Each plate was streaked by the quadrant method. The plates were incubated at 37C at 3:20pm.
6/02/21
6/02/21
The 2 plates were removed from the incubator and placed in Fridge B at 8:00am. pKK-mRuby3-TEV-1 has some good colonies, but they are less isolated. pKK-mRuby3-TEV-2 has multiple good colonies that are isolated. These plates were by HS on 6/01/21. They were inoculated and incubated for 16 hours before maxiprep.
6/03-05/21
6/03-05/21
Maxiprep was attempted on pKK-mRuby3-TEV1 and 2 on 6/03/21. Due to a shortage of solutions pKK-mRuby3-TEV1 and 2 were frozen after step 6 (the addition of PCI, then centrifuge, then transfer the top layer to a new tube). The Maxiprep was continued on 6/04/21. No pellets formed after the 100% ethanol step (step 8), so 1/10th of the volume of sodium acetate buffer was added. The tubes were then inverted 1 times and left overnight. On 6/05/21 the tubes were spun at 13,000g for 5 minutes where a pellet formed. Then the 40% ethanol step (step 9) was completed and the tubes were left for 2 nights to dry (step 10).
6/06/21
6/06/21
pKK-mRuby3-TEV1 and 2 (alpha) were resuspended in 1mL of water and left in Freezer A. Gel Electrophoresis was ran on these maxiprep sample. On lanes 2 and 3. They were at a very low concentration compared to what maxipreps normally produce. We will sequence to see if the DNA is correct.
6/07/21
6/07/21
pKK-mRuby3-TEV was streaked and plated twice by HS on 6/07/21. Each plate was streaked with the quadrant method. LB +Ampicillin plates were made on 6/01/21 by LC. Plates were incubated for 1 hour before streaking and isolation.
This plate indicates that pKK-mRuby3-TEV-1 was obtained from the bacterial stab and transformed successfully. However, streaking to obtain individual colonies was not as successful with this plate, as multiple colonies are grouped close together or are overlapping.
This plate indicates that pKK-mRuby3-TEV-2 was obtained from the bacterial stab and transformed successfully. Streaking was also successful, as there are several colonies that are of the right shape and size, and are separate from other colonies. This plate will likely be used for future maxi-preps.
6/08/21
6/08/21
Plates (pKK-mRuby3-TEV-1 and 2) were removed from the incubator by HS at 8am and placed in Fridge B.
Maxiprep was performed on pKK-mRuby3-TEV-1 and 2 (beta) because it was the second time the same colonies were inoculated. The maxiprep was done from the plates made on 6/02/21 by HS. Maxiprep was successful and the tubes were left overnight to dry.
6/09/21
6/09/21
pKK-mRuby3-TEV1 and 2 (beta) were resuspended in 0.5mL MBG Water and placed in Freezer C.
6/10/21
6/10/21
Gel Electrophoresis was ran on the second maxiprep of mRuby3. Lanes 2 and 3.
Page updated
Report abuse