Team:UNIPV-Pavia/Calendar/July

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Inoculum of <partinfo>BBa_J72007</partinfo> and <partinfo>BBa_J72013</partinfo> from LB agar plates.A glycerol stock was prepared for each culture.<br>
Inoculum of <partinfo>BBa_J72007</partinfo> and <partinfo>BBa_J72013</partinfo> from LB agar plates.A glycerol stock was prepared for each culture.<br>
Inoculum of <partinfo>BBa_J72008</partinfo> from LB agar plate grown at 30°C 220 rpm overnight.  
Inoculum of <partinfo>BBa_J72008</partinfo> from LB agar plate grown at 30°C 220 rpm overnight.  
 +
</td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana1 #July, 3rd|July, 3rd]]</td><td align="left">
 +
21° Bio-Lab - Glycerol stock for <partinfo>BBa_J72008</partinfo> was prepared.
</td></tr>
</td></tr>
<tr><th align="center" width="12%" colspan="2"><b>JULY: WEEK 2</b></th></tr>
<tr><th align="center" width="12%" colspan="2"><b>JULY: WEEK 2</b></th></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana2 #July, 5th|July, 5th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana2 #July, 5th|July, 5th]]</td><td align="left">
-
21° Bio-Lab - LB agar plates prepared.
+
22° Bio-Lab - LB agar plates prepared.
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana2 #July, 6th|July, 6th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana2 #July, 6th|July, 6th]]</td><td align="left">
-
22° Bio-Lab - We decided to perform a TECAN test to evaluate the strength of some promoters belonging to the Anderson Promoters Collection.
+
23° Bio-Lab - We decided to perform a TECAN test to evaluate the strength of some promoters belonging to the Anderson Promoters Collection.
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana2 #July, 7th|July, 7th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana2 #July, 7th|July, 7th]]</td><td align="left">
-
23° Bio-Lab -Competent cells preparation for:
+
24° Bio-Lab -Competent cells preparation for:
* MG1655
* MG1655
* BW53474
* BW53474
Line 67: Line 70:
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana2 #July, 8th|July, 8th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana2 #July, 8th|July, 8th]]</td><td align="left">
-
24° Bio-Lab - Transformation of <partinfo>BBa_J23118</partinfo> in our home-made competent strains  
+
25° Bio-Lab - Transformation of <partinfo>BBa_J23118</partinfo> in our home-made competent strains  
* MG1655
* MG1655
* BW25141
* BW25141
Line 73: Line 76:
* BW53474
* BW53474
*DH5alpha (as control)
*DH5alpha (as control)
-
to check transformation efficiency.</td></tr>
+
to check transformation efficiency.
 +
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana2 #July, 9th|July, 9th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana2 #July, 9th|July, 9th]]</td><td align="left">
-
25° Bio-Lab - All plates showed red colonies, so we calculated the efficiency of the strains.</td></tr>
+
26° Bio-Lab - All plates showed red colonies, so we calculated the efficiency of the strains.
 +
</td></tr>
<tr><th align="center" width="12%" colspan="2"><b>JULY: WEEK 3</b></th></tr>
<tr><th align="center" width="12%" colspan="2"><b>JULY: WEEK 3</b></th></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana3 #July, 12th|July, 12th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana3 #July, 12th|July, 12th]]</td><td align="left">
-
26° Bio-Lab -  
+
27° Bio-Lab - Phasins PhaP1 and PhaP2 were amplified by PCR and the results were sequenced .
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana3 #July, 13th|July, 13th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana3 #July, 13th|July, 13th]]</td><td align="left">
-
27° Bio-Lab -  
+
28° Bio-Lab - Phasins samples were sent to be sequenced. MiniPrep was performed. Ligations (from I11 till I19 and I74c5, I84c5, I104c5) were all performed
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana3 #July, 14th|July, 14th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana3 #July, 14th|July, 14th]]</td><td align="left">
-
28° Bio-Lab -  
+
29° Bio-Lab - PBHR68 plate showed colonies!!transformation of ligation of yesterday. Screening of <partinfo>BBa_J72007</partinfo> (CRIM), <partinfo>BBa_J72008</partinfo> (helper), pAH123 (helper) ans pCP20 (helper).
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana3 #July, 15th|July, 15th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana3 #July, 15th|July, 15th]]</td><td align="left">
-
29° Bio-Lab -  
+
30° Bio-Lab -Agar plates after transformation were checked(2 colonies were picked from every plate).
 +
MiniPrep was performed for:
 +
* PBHR68 (BioPlastic operon) -> 272 ng/ul
 +
* BBa_J72007(CRIM plasmid) -> 41 ng/ul (not clean spectrum at 230nm)
 +
* BBa_J72013(CRIM plasmid) -> 16 ng/ul 
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana3 #July, 16th|July, 16th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana3 #July, 16th|July, 16th]]</td><td align="left">
-
30° Bio-Lab -  
+
31° Bio-Lab - MiniPrep was performed on 23 falcon tubes containing I11 -> I19 and I7/8/10-4C5 ligations.
 +
We decided to performe a TECAN test  on our parts, in order to see if RFP was correctly excided from BBa_J231xx vector, since length of RFP and of our insert (RBS-luxI-tt) were similar.
</td></tr>
</td></tr>
<tr><th align="center" width="12%" colspan="2"><b>JULY: WEEK 4</b></th></tr>
<tr><th align="center" width="12%" colspan="2"><b>JULY: WEEK 4</b></th></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 19th|July, 19th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 19th|July, 19th]]</td><td align="left">
-
31° Bio-Lab -  
+
32° Bio-Lab - TECAN test showed that no RFP was produced from our parts.I14-1, I16-1, I17-1, I18-1, I19-1, I74c5-2, I84c5-2 and I12-2 were sequenced.
-
</td></tr>
+
Trasformation of RING into:
 +
* BW25141 (pir+)
 +
* BW25142 (pir116)
 +
* BW23474 (pir116)
 +
* DH5alpha
 +
* MG1655
 +
</td></tr>  
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 20th|July, 20th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 20th|July, 20th]]</td><td align="left">
-
32° Bio-Lab -  
+
33° Bio-Lab - Agar plates after transformation were checked (Single colonies were picked from plates).
 +
MiniPrep was performed on I10-1, I12-2, I3-1, I14-1, I17-1 and <partinfo>BBa_J23110</partinfo>.
 +
Following ligations were performed: I15_new, I14_4C5, I16_4C5, I17_4C5, I18_4C5 and I19_4C5.
 +
We incoulated from glycerol stock (3ul in 2ml LB+Amp):
 +
<partinfo>BBa_J23110</partinfo>, <partinfo>BBa_J23118</partinfo>, <partinfo>BBa_J23116</partinfo>, <partinfo>BBa_J23114</partinfo>, <partinfo>BBa_J23106</partinfo>, <partinfo>BBa_J23105</partinfo>, <partinfo>BBa_J23101</partinfo>, <partinfo>BBa_J23100</partinfo>, <partinfo>BBa_B0033</partinfo>
 +
in order to perform a TECAN test tomorrow.
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 21st|July, 21st]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 21st|July, 21st]]</td><td align="left">
-
33° Bio-Lab -
+
34° Bio-Lab -Tecan Test. This morning all the seven cultures were grown, For those cultures glycerol stocks were prepared and stored at -80°C. (Remaning 5 ml were used to perform MiniPrep). 
 +
Competent cells for cultures inoculated yesterday were prepared. Trasformations of ligations of yesterday were performed.
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 22nd|July, 22nd]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 22nd|July, 22nd]]</td><td align="left">
-
34° Bio-Lab -  
+
35° Bio-Lab - Plates were checked.
 +
Following ligations were performed: I17_4C5, I18_4C5, I19_4C5, I10-4C5, I12-4C5.
 +
Competent T9002 and MC1061 cells were transformed with a negative control in pSB4C5.
 +
We also performed a TECAN test in order to evaluate the ranking of sternght of promoters we inoculaed yesterday.
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 23rd|July, 23rd]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 23rd|July, 23rd]]</td><td align="left">
-
35° Bio-Lab -  
+
36° Bio-Lab - All plates incubated yesterday (T9002-4C5, MC1051-4C5 and PBHR68-4C5) showed colonies!
 +
Trasformations of ligations of yesterday were performed.
 +
We also received sequencing results (all correct).
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 24th|July, 24th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 24th|July, 24th]]</td><td align="left">
-
36° Bio-Lab -  
+
37° Bio-Lab - Two colonies from each plate incubated yesterday were picked.
 +
Colonies were counted to evaluate efficiency of transformation for our home made competent cells.
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 25th|July, 25th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana4 #July, 25th|July, 25th]]</td><td align="left">
-
37° Bio-Lab -  
+
38° Bio-Lab - Screening digestion was performed for colonies picked yesterday. MiniPrep and gel run was performed.
</td></tr>
</td></tr>
<tr><th align="center" width="12%" colspan="2"><b>JULY: WEEK 5</b></th></tr>
<tr><th align="center" width="12%" colspan="2"><b>JULY: WEEK 5</b></th></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #July, 26th|July, 26th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #July, 26th|July, 26th]]</td><td align="left">
-
38° Bio-Lab - <br>  
+
39° Bio-Lab - Ligation of I15_4C5. <br>  
12° Meeting - Updating about Bio-Lab activity and organization of our trip to Boston!  
12° Meeting - Updating about Bio-Lab activity and organization of our trip to Boston!  
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #July, 27th|July, 27th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #July, 27th|July, 27th]]</td><td align="left">
-
39° Bio-Lab -  
+
40° Bio-Lab - Sample of I15-1 was prepared for sequencing.I14_4C5, I16_4C5, I17_4C5, I18_4C5 and I19_4C5 were cotrasformed in T9002.
 +
MiniPrep was performed on MC1, MC2.
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #July, 28th|July, 28th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #July, 28th|July, 28th]]</td><td align="left">
-
40° Bio-Lab -  
+
41° Bio-Lab - Today we diluted primers to modify PhaPs and performed a PCR with them in order to mutagenize this BioBrick to get its prefix Standard/Silver compliant and suffix Silver compliant.
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #July, 29th|July, 29th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #July, 29th|July, 29th]]</td><td align="left">
-
41° Bio-Lab -  
+
42° Bio-Lab - PCR was prepared in order to amplify Phasins Phap (<partinfo>BBa_K208001</partinfo>).After that phasins were gel extracted and digested.
 +
A tecan test was performed.
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #July, 30th|July, 30th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #July, 30th|July, 30th]]</td><td align="left">
-
42° Bio-Lab -  
+
43° Bio-Lab - Agar plates were checked and efficiency was calculated.
 +
Miniprep of <partinfo>BBa_J04450</partinfo> was performed.
 +
Ligation of I20, I21.
</td></tr>
</td></tr>
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #July, 31th|July, 31th]]</td><td align="left">
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #July, 31th|July, 31th]]</td><td align="left">
-
43° Bio-Lab -  
+
44° Bio-Lab - Transformation of I20 and I21 into E. coli DH5-alpha.
</td></tr>
</td></tr>
</table>
</table>

Latest revision as of 12:49, 22 October 2010


JULY



Bio-Lab activity proceeds!

Select the day you are interested in to find out the details!

DATEACTIVITY
July, 1st

19° Bio-Lab - I7-3,4 and 5 and I8-3, 4 and 5 were prepared for TECAN test.
Of the plates incubated yesterday, a colony was picked and grown in 1ml LB (+ antibiotic) for 6 hours at 37°C 220 rpm. Other parts were received from Anderson Lab.

July, 2nd

20° Bio-Lab - TECAN test provided encouraging results, showing that I7-3, I7-5, I8-4 and I8-5 produced GFP.
Other culture (I7-1, I7-2, I7-4, I8-1, I8-2, I8-3) did not produce GFP, so they were thrown away. A furhter screening were performed on I7-3, I7-5, I8-4, I8-5 and this time all the clones are OK . Inoculum of <partinfo>BBa_J72007</partinfo> and <partinfo>BBa_J72013</partinfo> from LB agar plates.A glycerol stock was prepared for each culture.
Inoculum of <partinfo>BBa_J72008</partinfo> from LB agar plate grown at 30°C 220 rpm overnight.

July, 3rd

21° Bio-Lab - Glycerol stock for <partinfo>BBa_J72008</partinfo> was prepared.

JULY: WEEK 2
July, 5th

22° Bio-Lab - LB agar plates prepared.

July, 6th

23° Bio-Lab - We decided to perform a TECAN test to evaluate the strength of some promoters belonging to the Anderson Promoters Collection.

July, 7th

24° Bio-Lab -Competent cells preparation for:

  • MG1655
  • BW53474
  • BW25141
  • BW25142

PCR was performed to amplify Phasins.
11° Meeting - Updating about Bio-Lab activity.

July, 8th

25° Bio-Lab - Transformation of <partinfo>BBa_J23118</partinfo> in our home-made competent strains

  • MG1655
  • BW25141
  • BW25142
  • BW53474
  • DH5alpha (as control)

to check transformation efficiency.

July, 9th

26° Bio-Lab - All plates showed red colonies, so we calculated the efficiency of the strains.

JULY: WEEK 3
July, 12th

27° Bio-Lab - Phasins PhaP1 and PhaP2 were amplified by PCR and the results were sequenced .

July, 13th

28° Bio-Lab - Phasins samples were sent to be sequenced. MiniPrep was performed. Ligations (from I11 till I19 and I74c5, I84c5, I104c5) were all performed

July, 14th

29° Bio-Lab - PBHR68 plate showed colonies!!transformation of ligation of yesterday. Screening of <partinfo>BBa_J72007</partinfo> (CRIM), <partinfo>BBa_J72008</partinfo> (helper), pAH123 (helper) ans pCP20 (helper).

July, 15th

30° Bio-Lab -Agar plates after transformation were checked(2 colonies were picked from every plate). MiniPrep was performed for:

  • PBHR68 (BioPlastic operon) -> 272 ng/ul
  • BBa_J72007(CRIM plasmid) -> 41 ng/ul (not clean spectrum at 230nm)
  • BBa_J72013(CRIM plasmid) -> 16 ng/ul
July, 16th

31° Bio-Lab - MiniPrep was performed on 23 falcon tubes containing I11 -> I19 and I7/8/10-4C5 ligations. We decided to performe a TECAN test on our parts, in order to see if RFP was correctly excided from BBa_J231xx vector, since length of RFP and of our insert (RBS-luxI-tt) were similar.

JULY: WEEK 4
July, 19th

32° Bio-Lab - TECAN test showed that no RFP was produced from our parts.I14-1, I16-1, I17-1, I18-1, I19-1, I74c5-2, I84c5-2 and I12-2 were sequenced. Trasformation of RING into:

  • BW25141 (pir+)
  • BW25142 (pir116)
  • BW23474 (pir116)
  • DH5alpha
  • MG1655
July, 20th

33° Bio-Lab - Agar plates after transformation were checked (Single colonies were picked from plates). MiniPrep was performed on I10-1, I12-2, I3-1, I14-1, I17-1 and <partinfo>BBa_J23110</partinfo>. Following ligations were performed: I15_new, I14_4C5, I16_4C5, I17_4C5, I18_4C5 and I19_4C5. We incoulated from glycerol stock (3ul in 2ml LB+Amp): <partinfo>BBa_J23110</partinfo>, <partinfo>BBa_J23118</partinfo>, <partinfo>BBa_J23116</partinfo>, <partinfo>BBa_J23114</partinfo>, <partinfo>BBa_J23106</partinfo>, <partinfo>BBa_J23105</partinfo>, <partinfo>BBa_J23101</partinfo>, <partinfo>BBa_J23100</partinfo>, <partinfo>BBa_B0033</partinfo> in order to perform a TECAN test tomorrow.

July, 21st

34° Bio-Lab -Tecan Test. This morning all the seven cultures were grown, For those cultures glycerol stocks were prepared and stored at -80°C. (Remaning 5 ml were used to perform MiniPrep). Competent cells for cultures inoculated yesterday were prepared. Trasformations of ligations of yesterday were performed.

July, 22nd

35° Bio-Lab - Plates were checked. Following ligations were performed: I17_4C5, I18_4C5, I19_4C5, I10-4C5, I12-4C5. Competent T9002 and MC1061 cells were transformed with a negative control in pSB4C5. We also performed a TECAN test in order to evaluate the ranking of sternght of promoters we inoculaed yesterday.

July, 23rd

36° Bio-Lab - All plates incubated yesterday (T9002-4C5, MC1051-4C5 and PBHR68-4C5) showed colonies! Trasformations of ligations of yesterday were performed. We also received sequencing results (all correct).

July, 24th

37° Bio-Lab - Two colonies from each plate incubated yesterday were picked. Colonies were counted to evaluate efficiency of transformation for our home made competent cells.

July, 25th

38° Bio-Lab - Screening digestion was performed for colonies picked yesterday. MiniPrep and gel run was performed.

JULY: WEEK 5
July, 26th

39° Bio-Lab - Ligation of I15_4C5.
12° Meeting - Updating about Bio-Lab activity and organization of our trip to Boston!

July, 27th

40° Bio-Lab - Sample of I15-1 was prepared for sequencing.I14_4C5, I16_4C5, I17_4C5, I18_4C5 and I19_4C5 were cotrasformed in T9002. MiniPrep was performed on MC1, MC2.

July, 28th

41° Bio-Lab - Today we diluted primers to modify PhaPs and performed a PCR with them in order to mutagenize this BioBrick to get its prefix Standard/Silver compliant and suffix Silver compliant.

July, 29th

42° Bio-Lab - PCR was prepared in order to amplify Phasins Phap (<partinfo>BBa_K208001</partinfo>).After that phasins were gel extracted and digested. A tecan test was performed.

July, 30th

43° Bio-Lab - Agar plates were checked and efficiency was calculated. Miniprep of <partinfo>BBa_J04450</partinfo> was performed. Ligation of I20, I21.

July, 31th

44° Bio-Lab - Transformation of I20 and I21 into E. coli DH5-alpha.