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JULY
Bio-Lab activity proceeds!
Select the day you are interested in to find out the details!
| DATE | ACTIVITY |
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| 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.
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| 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.
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| JULY: WEEK 2 |
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| July, 5th |
22° Bio-Lab - LB agar plates prepared.
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| 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.
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| JULY: WEEK 3 |
|---|
| July, 12th |
27° Bio-Lab - Phasins PhaP1 and PhaP2 were amplified by PCR and the results were sequenced .
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| 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.
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| JULY: WEEK 4 |
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| 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.
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| 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).
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| 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.
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| July, 25th |
38° Bio-Lab - Screening digestion was performed for colonies picked yesterday. MiniPrep and gel run was performed.
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| 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!
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| 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.
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| 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.
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| 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.
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| July, 31th |
44° Bio-Lab - Transformation of I20 and I21 into E. coli DH5-alpha.
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