Team:BYU Provo/Notebook/Cholera - Enzyme/July-August/Period3/Dailylog

From 2013.igem.org

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(Created page with "{{TeamBYUProvo}} <br> {| width="100%" | colspan="3" | <font color="#333399" size="5" font face="Calibri"> '''Cholera - Enzymes Notebook: May 1 - May 14 Daily Log'''</font> <br...")
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<font size="4">'''8/12/13'''</font>
<font size="4">'''8/12/13'''</font>
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We ran DspB and Savinase on gel but the results were not promising. There were two very faint bands for DspB, but not enough for us to use. Savinase did not have any bands showing at all. We then reran the Phusion PCR for our samples following the protocol listed in the Grose lab protocol packet.
 +
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Our samples were:
 +
#DspB
 +
#DspB Control
 +
#Savinase
 +
#Savinase Control
 +
 +
 +
We also reseeded new cholera overnights by adding 4 mL of SLB and 50 uL of our previous cholera overnights.
 +
 +
<br>
 +
 +
<font size="4">'''8/13/13'''</font>
 +
 +
We set up PCR again for DspB and Savinase. The PCR tubes were labeled:
 +
#DspB
 +
#Savinase
 +
 +
<br>
 +
 +
<font size="4">'''8/14/13'''</font>
 +
 +
We ran the PCR products from yesterday on gel, but nothing showed, not even the ladder. This indicates that when the gel was prepared, the ethidium bromide was not added as it was supposed to be.
 +
 +
We made a new gel and re-ran our PCR products:
 +
#Ladder
 +
#DspB
 +
#DspB Control
 +
#Savinase
 +
#Savinase Control
 +
 +
[IMAGE]
 +
 +
There was a faint band for the DspB, but nothing for Savinase. We will try to move forward with our DspB, but may need to restreak the Bacillis subtilis for our Savinase enzyme.
 +
 +
<br>
 +
 +
<font size="4">'''8/21/13'''</font>
Insert Text
Insert Text

Revision as of 20:26, 6 September 2013


Cholera - Enzymes Notebook: May 1 - May 14 Daily Log



Overview
March-April
May-June
July-August
September-October

8/5/13

We got a frozen culture of V. cholerae from Dr. Robison’s lab. Cells are in 20% glycerol. We are waiting for the DspB colony PCR to finish so that we can grow up both the DspB and Savinase at the same time to sequence them and get them ready for protein purification.


8/7/13

The DspB colony PCR didn’t work. We re-ran the PCR yesterday, so we will run the product on gel today, clean up the PCR and sequence it. We also prepped the Savinase for sequencing today since the DspB didn’t work and we don’t want to wait any longer. We also streaked a new plate of cholera with the culture that we received from Dr. Robison’s lab. New plate was placed in the 37° incubator.


8/9/13

Phusion PCR of Savinase and DspB. We ran two samples of each with a control:

  1. DspB
  2. DspB
  3. DspB Control
  4. Savinase
  5. Savinase
  6. Savinase Control


8/12/13

We ran DspB and Savinase on gel but the results were not promising. There were two very faint bands for DspB, but not enough for us to use. Savinase did not have any bands showing at all. We then reran the Phusion PCR for our samples following the protocol listed in the Grose lab protocol packet.

Our samples were:

  1. DspB
  2. DspB Control
  3. Savinase
  4. Savinase Control


We also reseeded new cholera overnights by adding 4 mL of SLB and 50 uL of our previous cholera overnights.


8/13/13

We set up PCR again for DspB and Savinase. The PCR tubes were labeled:

  1. DspB
  2. Savinase


8/14/13

We ran the PCR products from yesterday on gel, but nothing showed, not even the ladder. This indicates that when the gel was prepared, the ethidium bromide was not added as it was supposed to be.

We made a new gel and re-ran our PCR products:

  1. Ladder
  2. DspB
  3. DspB Control
  4. Savinase
  5. Savinase Control

[IMAGE]

There was a faint band for the DspB, but nothing for Savinase. We will try to move forward with our DspB, but may need to restreak the Bacillis subtilis for our Savinase enzyme.


8/21/13

Insert Text


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