Team:Valencia/Notebook/October

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(Difference between revisions)

Latest revision as of 22:14, 29 October 2008

October 1st

- We repeated the previous experiment:
      Four strains in LCCs with SP medium.
      1 % Galactose induction at the beginning. Initial O.D. 2
      270 rpm.
       Initial temperature 28 ºC.

October 2nd

- Team meeting: We talked about the biobricks and some experiments left.

October 7th

- We repeated the previous experiment:
      Four strains in LCCs with SP medium.
      1 % Galactose induction at the beginning. Initial O.D. 2
      270 rpm.
       Initial temperature 28 ºC.

October 13th

- Total DNA was extracted from our yeast strains
- UCP-1 was amplified by PCR using oligonucleotides matching the sequence and bearing the appropriate Biobrick prefix and suffix.

October 14th

- We performed an agarose gel electrophoresis in order to check our PCR, the results was: Intense amplicons, of the expected size, (about 1 kb) were produced for UCP-1, 175-deleted and 76 deleted.

October 17th

- Team meeting: We discussed several things that need to be finished, such as sending DNA, preparing presentation, poster, wiki...

October 20th

- Preparing vectors, competent cells were transformed with: pSB1AK3 and pUC18 plasmids.

October 23rd

- Plasmids were extracted (High pure miniprep plasmid isolation kit ROCHE).

- Plasmids were digested with EcoRI and PstI

October 24th

- Ligating Biobricks into plasmids: Inserts were run into agarose gels and DNA bands excised with a clean scalpel. DNA was extracted from agarose blocks (ultra clean gel spin, DNA purification Kit, MO BIO laboratories). T4 Ligase was used to ligate inserts and vectors.

- Competent cells were transformed and resulting colonies (Amp LB) screened with Fw and Rv primers to confirm the presence of inserts.

October 27th

- pSB1AK3 containing UCP-1, 175-deleted and 76-deleted were sent to the Registry.

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+| <Font Color="#c0c0c0">29</Font> || <Font Color="#c0c0c0">30</Font> || &nbsp;&nbsp;[[Team:Valencia/Notebook/October#October 1st | 1]]|| &nbsp;[[Team:Valencia/Notebook/October#October 2nd | 2]] || &nbsp;3 || &nbsp;4 || &nbsp;5
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-| &nbsp;6 || &nbsp;7 || &nbsp;&nbsp;8|| 9 || 10 || 11|| 12
+| &nbsp;6 || &nbsp;[[Team:Valencia/Notebook/October#October 7th |7]]  || &nbsp;&nbsp;8|| 9 || 10 || 11|| 12
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-| 13 || 14 || &nbsp;15 || 16 || 17 || 18 || 19
+| [[Team:Valencia/Notebook/October#October 13th |13]] || [[Team:Valencia/Notebook/October#October 14th |14]]  || &nbsp;15 || 16 || [[Team:Valencia/Notebook/October#October 17th |17]] || 18 || 19
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-| 20 || 21 || &nbsp;22 || 23 || 24 || 25 || 26
+| [[Team:Valencia/Notebook/October#October 20th |20]] || 21 || &nbsp;22 || [[Team:Valencia/Notebook/October#October 23rd |23]]  || [[Team:Valencia/Notebook/October#October 24th |24]]  || 25 || 26
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+| [[Team:Valencia/Notebook/October#October 27th |27]]  || 28 || &nbsp;29 || 30 || 31 || <Font Color="#c0c0c0">&nbsp;1</Font> || <Font Color="#c0c0c0">&nbsp;2</Font>
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 &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;270 rpm. <br>
 &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Initial temperature 28 ºC.<br>
+=== October 2nd ===
+- Team meeting: We talked about the biobricks and some experiments left.
+=== October 7th ===
+- We repeated the previous experiment:<br>
+&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Four strains in LCCs with [[Team:Valencia/Notebook/Protocols#SP liquid medium | SP medium]]. <br>
+&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;1 % Galactose induction at the beginning. Initial O.D. 2 <br>
+&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;270 rpm. <br>
+&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Initial temperature 28 ºC.<br>
+=== October 13th ===
+- Total DNA was extracted from our yeast strains <br>
+- UCP-1 was amplified by [[Team:Valencia/Parts#Preparing_inserts|PCR]] using oligonucleotides matching the sequence and bearing the appropriate Biobrick prefix and suffix.
+=== October 14th ===
+- We performed an [[Team:Valencia/Parts#Preparing_inserts|agarose gel electrophoresis]] in order to check our PCR, the results was: Intense amplicons, of the expected size, (about 1 kb) were produced for UCP-1, 175-deleted and 76 deleted.
+=== October 17th ===
+- Team meeting: We discussed several things that need to be finished, such as sending DNA, preparing presentation, poster, wiki...
+=== October 20th ===
+- Preparing vectors, competent cells were transformed with: pSB1AK3 and pUC18 plasmids.
+=== October 23rd ===
+- Plasmids were extracted (High pure miniprep plasmid isolation kit ROCHE).
+- Plasmids were digested with EcoRI and PstI
+=== October 24th ===
+- [[Team:Valencia/Parts#Ligating_Biobricks_into_plasmids|Ligating Biobricks]] into plasmids:
+Inserts were run into agarose gels and DNA bands excised with a clean scalpel. DNA was extracted from agarose blocks (ultra clean gel spin, DNA purification Kit, MO BIO laboratories). T4 Ligase was used to ligate inserts and vectors.
+- Competent cells were transformed and resulting colonies (Amp LB) screened with Fw and Rv primers to confirm the presence of inserts.
+=== October 27th ===
+- pSB1AK3 containing UCP-1, 175-deleted and 76-deleted were sent to the Registry.