Team:UNIPV-Pavia/Calendar/August/week1

From 2011.igem.org

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TOP10 competent cells, carrying <a href="http://partsregistry.org/wiki/index.php/Part:BBa_T9002">BBa_T9002</a> construct, were transformed with ENTERO4C5 to confer resistance to Cloramphenicol (12.5 mg/ml).
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TOP10 competent cells, harboring <a href="http://partsregistry.org/wiki/index.php/Part:BBa_T9002">BBa_T9002</a> construct, were co-transformed with ENTERO4C5 to confer resistance to Cloramphenicol (12.5 mg/ml).
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Revision as of 13:44, 16 August 2011

UNIPV TEAM 2011

March
M T W T F S S
  1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30 31    

April
M T W T F S S
        1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
25 26 27 28 29 30

May
M T W T F S S
            1
2 3 4 5 6 7 8
9 10 11 12 13 14 15
16 17 18 19 20 21 22
23 24 25 26 27 28 29
30 31

June
M T W T F S S
    1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30

July
M T W T F S S
        1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
25 26 27 28 29 30 31

August
M T W T F S S
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 30 31

September
M T W T F S S
      1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30

October
M T W T F S S
          1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31

AUGUST: WEEK 1

August, 1st

For each plate from the previous day, two colonies were picked.

Next, MGZ1 competent cells were prepared according to protocol. They were put in 22 100 μl test-tubes.

August, 2nd

Plasmids containing E28AGAIN-1, E28AGAIN-2, E41AGAIN-1, E28AGAIN-2, E37-2, E38-1. E39-1, E40-2 and E42-1 were extracted with MiniPrep kit from ON saturated cultures; purified DNA was quantified with NanoDrop Spectrophotometer:

Plasmid DNA (ng/μl)
E28AGAIN-1 21.4
E28AGAIN-2 21.0
E41AGAIN-1 33.7
E41AGAIN-2 27.7
E37-2 18.5
E38-1 26.0
E39-1 21.6
E40-2 19.9
E42-1 25.1

J101-31, J101-E5, J101-E7 were transformed in MGZ1 competent cells.

Digestions of E28AGAIN-1, E28AGAIN-2, E41AGAIN-1 and E42AGAIN-2 were performed for screening; a 5x mix was prepared and for each sample the volumes specified in the table below were used:

DNA (μl) H2O (μl) Enzyme 1 (μl) Enzyme 2 (μl) Buffer H (μl) Final Volume (μl)
5 16.5 0.5 EcoRI 0.5 Pstl 2.5 25

TOP10 competent cells, harboring BBa_T9002 construct, were co-transformed with ENTERO4C5 to confer resistance to Cloramphenicol (12.5 mg/ml).