Difference between revisions of "Part:BBa K1022114"
Line 21: Line 21:
 
Description :  
 
Description :  
 
The <i>E.coli</i> cells transformed with pT7 lysis cassette [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1022114  BBa_K1022114] is grown on a plate reader which is capable of shaking and heating to 37˚C to take readings of the cells in exponential phase at every 10 minutes. Different range of IPTG concentration is used to characterize the bio – brick  [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1022114  BBa_K1022114].  At time point, 160 minutes, IPTG is added according to the table below.  
 
The <i>E.coli</i> cells transformed with pT7 lysis cassette [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1022114  BBa_K1022114] is grown on a plate reader which is capable of shaking and heating to 37˚C to take readings of the cells in exponential phase at every 10 minutes. Different range of IPTG concentration is used to characterize the bio – brick  [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1022114  BBa_K1022114].  At time point, 160 minutes, IPTG is added according to the table below.  
 +
 +
                <table >
 +
                    <tr>
 +
                        <td></td>
 +
                        <td>
 +
                        No cells + 1mM
 +
                        </td>
 +
                        <td >0.1mM</td>
 +
                        <td>0.2m</td>
 +
                        <td>0.3mM</td>
 +
                        <td>0.4mM</td>
 +
                        <td>0.5mM</td>
 +
                        <td>0.6mM</td>
 +
                        <td>
 +
                          0.7mM
 +
                        </td>
 +
                        <td>
 +
                          0.8mM
 +
                        </td>
 +
                        <td>
 +
                          0.9mM
 +
                        </td>
 +
                        <td>
 +
                          1mM
 +
                        </td>
 +
                        <td>
 +
                          Cell + No IPTG
 +
                        </td>
 +
                    </tr>
 +
 +
    <tr>
 +
                        <td>
 +
                        LB(µL)
 +
                        </td>
 +
                        <td >
 +
                          90
 +
                        </td>
 +
                        <td>
 +
                            94
 +
                        </td>
 +
                        <td>93</td>
 +
                        <td>92</td>
 +
                                              <td>91</td>
 +
                        <td>90</td>
 +
                        <td>89</td>
 +
                        <td>88</td>
 +
                        <td>87</td>
 +
                        <td>86</td>
 +
                        <td>85</td>
 +
                        <td>95</td>     
 +
                    </tr>
 +
 +
<tr>
 +
                        <td>
 +
                        Cells(µL)
 +
                        </td>
 +
                        <td>-</td>   
 +
                        <td>5</td>
 +
                        <td>5</td>     
 +
                        <td>5</td>
 +
                        <td>5</td>
 +
                        <td>5</td> 
 +
                        <td>5</td> 
 +
                        <td>5</td>   
 +
                        <td>5</td>       
 +
                        <td>5</td> 
 +
                        <td>5</td>
 +
                        <td>5</td>               
 +
                     
 +
                    </tr>
 +
 +
<tr>
 +
                        <td>
 +
                        10X IPTG(µL)
 +
                        </td>
 +
                        <td>10</td>   
 +
                        <td>1</td>
 +
                        <td>2</td>     
 +
                        <td>3</td>
 +
                        <td>4</td> 
 +
                        <td>5</td>
 +
                        <td>6</td>
 +
                        <td>7</td>
 +
                        <td>8</td>
 +
                        <td>9</td> 
 +
                        <td>10</td>     
 +
                        <td>-</td>               
 +
                     
 +
                    </tr>
 +
 +
</table>
 +
 +
 +
  
 
[[Image:Lysis TU Delft.jpg|450px|center]]
 
[[Image:Lysis TU Delft.jpg|450px|center]]

Revision as of 09:51, 3 October 2013

pT7 : Lysis Device

This bio-brick codes for the promoter pT7(BBa_I712074) followed by the lysis device(BBa_K112808).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1404
    Illegal NheI site found at 1427
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1009
    Illegal AgeI site found at 1079
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1660


Characterization

For more info, visit [http://2013.igem.org/Team:TU-Delft/Killswitch TU Delft iGEM13 Wiki !!]

The construct is made by ligating the pT7 promoter BBa_I712074 in front of the lysis device from the biobrick BBa_K112808. This is analyze how the lysis device can be controlled if needed.

114.jpg

Description : The E.coli cells transformed with pT7 lysis cassette BBa_K1022114 is grown on a plate reader which is capable of shaking and heating to 37˚C to take readings of the cells in exponential phase at every 10 minutes. Different range of IPTG concentration is used to characterize the bio – brick BBa_K1022114. At time point, 160 minutes, IPTG is added according to the table below. 

                        No cells + 1mM
0.1mM 0.2m 0.3mM 0.4mM 0.5mM 0.6mM 
                         0.7mM

                         0.8mM

                         0.9mM

                         1mM

                         Cell + No IPTG

                        LB(µL)

                          90

                           94
93 92 91 90 89 88 87 86 85 95 
                        Cells(µL)
- 5 5 5 5 5 5 5 5 5 5 5 
                       10X IPTG(µL)
10 1 2 3 4 5 6 7 8 9 10 -



Lysis TU Delft.jpg