Team:UNIPV-Pavia/Calendar/August/settimana3

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(Difference between revisions)
(→August, 17th)
(→August, 18th)
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Glycerol stock was prepared for I154C5-T9002.
Glycerol stock was prepared for I154C5-T9002.
 +
 +
----
 +
The following cultures were miniprepped and quantified with Nanodrop:
 +
*I26: 67,5 ng/ul
 +
*<partinfo>BBa_K105012</partinfo>: 60,9 ng/ul
 +
*<partinfo>BBa_B0034</partinfo>: 64,5 ng/ul
 +
*<partinfo>BBa_F2620</partinfo>: 91,8 ng/ul
 +
*I20M-1: 191,8 ng/ul
 +
*I20A-1: 176,2 ng/ul
 +
*I21M-3: 156,2 ng/ul
 +
*I21-4: 76,3 ng/ul
 +
*I0: 121,2 ng/ul
 +
*INTEIN: 143,9 ng/ul
 +
<partinfo>J13002</partinfo>: 31,9 ng/ul
 +
 +
and than digested as follows:
 +
{| border="1" align='center'
 +
|  ''Culture''    ||  ''Kind''  ||  ''Final reaction volume (ul) '' ||  ''DNA (ul)''  ||  ''H20 (ul)''  ||  ''Enzyme 1''  ||  ''Enzyme 2'' ||  ''Buffer H''
 +
|-
 +
|    I26  ||  Insert || 25 ||  21,5  ||  0  ||    0,5 EcoRI  || 0,5 SpeI  || 2,5
 +
|-
 +
|    <partinfo>BBa_K105012</partinfo>  ||  Vector || 25 ||  16,5  ||  5  ||      0,5 E  || 0,5 S  || 2,5
 +
|-
 +
|    <partinfo>BB_B0034</partinfo>  || Vector || 25 ||  15,5  || 6  ||    0,5 E  || 0,5 S    || 2,5
 +
|-
 +
|  <partinfo>BB_F2620</partinfo>  ||  Insert || 25 ||  16,5  ||  5  || 0,5 E  || 0,5 S  || 2,5
 +
|-
 +
|    I20M-1  ||  Insert/Screening || 25 ||  8  ||  13,5  ||  0,5 E  || 0,5 S  || 2,5
 +
|-
 +
|    I20M-1  ||  Insert/Screening || 25 ||  8  ||  13,5  ||  0,5 X || 0,5 PstI  || 2,5
 +
|-
 +
|    I20A-1  ||  Insert/Screening || 25 ||  13  ||  8,5  ||  0,5 E  || 0,5 S  || 2,5
 +
|-
 +
|    I20A-1  ||  Insert/Screening || 25 ||  13  ||  8,5  ||  0,5 X  || 0,5 P  || 2,5
 +
|-
 +
|    I21-4  ||  Insert/Screening || 25 ||  13  ||  8,5  ||  0,5 E  || 0,5 S  || 2,5
 +
|-
 +
|    I21-4  ||  Vector || 25 ||  13,1  ||  8,4  ||  0,5 E  || 0,5 X  || 2,5
 +
|-
 +
|    I0  ||  Vector || 25 ||  8,2  ||  13,3  || 0,5 E  || 0,5 XbaI  || 2,5
 +
|-
 +
|    INTEIN  ||  Insert || 25 ||  10,5  ||  11  || 0,5 E  || 0,5 S  || 2,5
 +
|-
 +
|  <partinfo>BB_J13002</partinfo>  ||  Vector || 25 ||  21,5  ||  0  ||  0,5 S  || 0,5 P  || 2,5
 +
|-
 +
|}
==August, 19th==
==August, 19th==

Revision as of 10:54, 20 August 2010
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AUGUST: WEEK 3


August, 16th

Resuspension from iGEM 2010 Distribution Kit of <partinfo>BBa_J13002</partinfo> (Plate 1, Well 13B) and <partinfo>BBa_I13522</partinfo> (Plate 2, Well 8A).

Transformation (1 ul) of <partinfo>BBa_J13002</partinfo>, <partinfo>BBa_I13522</partinfo>, INTEIN, CREAM (Mr.Gene) into 100 ul of E. coli TOP10 home-made competent cells.

Transformation of 15-4C5 (1 ul) into in re-competentized E. coli TOP10 carrying <partinfo>BBa_T9002</partinfo>.

Transfomants were plated on LB+Amp 100 ug/ml agar plates, the last one on LB+Amp100+Cm12,5 agar plates and were let grow ON at 37°C.

August, 17th

All plates showed colonies.

Pick of <partinfo>BBa_J13002</partinfo>, <partinfo>BBa_I13522</partinfo>, INTEIN, CREAM, co-trasformed <partinfo>BBa_T9002</partinfo> and inoculum into 1 ml of LB+Amp, LB+Kan, LB+Cm, respectively, to make glycerol stocks. Falcons were than refilled for the minipreps of the following day.

Inoculum of I5, I26, <partinfo>BBa_K105012</partinfo>, <partinfo>BBa_B0034</partinfo>, I20M-1, I20A-1, I21M-3, I21-4, I21A-7, I0 for minipreps of the following day.

Cultures were diluted (5ul in 2ml LB+antibiotic) for TECAN test:

I7-A I8-5 D I9-1
I10-B I12-2 A2
<partinfo>BBa_B0032</partinfo> I144C5-T9002 I164C5-T9002
I174C5-T9002 I184C5-T9002 I194C5-T9002
ENTERO4C5-T9002

Glycerol stock was prepared for ENTERO4C5-T9002.

August, 18th

Cultures were diluted (5ul in 2ml LB+antibiotic) for TECAN test:

I7-A I8-5 D I9-1
I10-B I12-2 A2
<partinfo>BBa_B0032</partinfo> I144C5-T9002 I154C5-T9002
I164C5-T9002 I174C5-T9002 I184C5-T9002
I194C5-T9002 ENTERO4C5-T9002

Glycerol stock was prepared for I154C5-T9002.

The following cultures were miniprepped and quantified with Nanodrop:

  • I26: 67,5 ng/ul
  • <partinfo>BBa_K105012</partinfo>: 60,9 ng/ul
  • <partinfo>BBa_B0034</partinfo>: 64,5 ng/ul
  • <partinfo>BBa_F2620</partinfo>: 91,8 ng/ul
  • I20M-1: 191,8 ng/ul
  • I20A-1: 176,2 ng/ul
  • I21M-3: 156,2 ng/ul
  • I21-4: 76,3 ng/ul
  • I0: 121,2 ng/ul
  • INTEIN: 143,9 ng/ul

<partinfo>J13002</partinfo>: 31,9 ng/ul

and than digested as follows:

Culture Kind Final reaction volume (ul) DNA (ul) H20 (ul) Enzyme 1 Enzyme 2 Buffer H
I26 Insert 25 21,5 0 0,5 EcoRI 0,5 SpeI 2,5
<partinfo>BBa_K105012</partinfo> Vector 25 16,5 5 0,5 E 0,5 S 2,5
<partinfo>BB_B0034</partinfo> Vector 25 15,5 6 0,5 E 0,5 S 2,5
<partinfo>BB_F2620</partinfo> Insert 25 16,5 5 0,5 E 0,5 S 2,5
I20M-1 Insert/Screening 25 8 13,5 0,5 E 0,5 S 2,5
I20M-1 Insert/Screening 25 8 13,5 0,5 X 0,5 PstI 2,5
I20A-1 Insert/Screening 25 13 8,5 0,5 E 0,5 S 2,5
I20A-1 Insert/Screening 25 13 8,5 0,5 X 0,5 P 2,5
I21-4 Insert/Screening 25 13 8,5 0,5 E 0,5 S 2,5
I21-4 Vector 25 13,1 8,4 0,5 E 0,5 X 2,5
I0 Vector 25 8,2 13,3 0,5 E 0,5 XbaI 2,5
INTEIN Insert 25 10,5 11 0,5 E 0,5 S 2,5
<partinfo>BB_J13002</partinfo> Vector 25 21,5 0 0,5 S 0,5 P 2,5

August, 19th

August, 20th

August, 21st

August, 22nd



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