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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1404
Illegal NheI site found at 1427 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1009
Illegal AgeI site found at 1079 - 1000INCOMPATIBLE 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.
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 | - |