Difference between revisions of "Team:Pasteur Paris/Microbiology week10"
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1. Realize a Mastermix and store it on ice : <br /> | 1. Realize a Mastermix and store it on ice : <br /> | ||
<table> | <table> | ||
| − | <caption align="bottom" align="center">Table | + | <caption align="bottom" align="center"><i><p> <U>Table 113</U></p></i></caption> |
<thead> | <thead> | ||
<tr> | <tr> | ||
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<h6><U>Results :</U></h6><br /><br /> | <h6><U>Results :</U></h6><br /><br /> | ||
<center><img src = "https://static.igem.org/mediawiki/2016/c/cf/Week_10_152Electrophoresis_with_the_results_of_digestion.jpg"; alt "Gel of the results of digestion"/> | <center><img src = "https://static.igem.org/mediawiki/2016/c/cf/Week_10_152Electrophoresis_with_the_results_of_digestion.jpg"; alt "Gel of the results of digestion"/> | ||
| − | <i><p> <U>Figure | + | <i><p> <U>Figure 18 :</U> Gel of the results of digestion </i></p></center><br /> |
<br /><br /><br /> | <br /><br /><br /> | ||
</p> | </p> | ||
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1. Prepare the following tubes :<br /> | 1. Prepare the following tubes :<br /> | ||
<table> | <table> | ||
| − | <caption align="bottom" align="center">Table | + | <caption align="bottom" align="center"><i><p> <U>Table 114</U></p></i></caption> |
<thead> | <thead> | ||
<tr> | <tr> | ||
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3. Add 1 µl of DNA following the number of tubes : <br /><br /> | 3. Add 1 µl of DNA following the number of tubes : <br /><br /> | ||
<table> | <table> | ||
| − | <caption align="bottom" align="center">Table | + | <caption align="bottom" align="center"><i><p> <U>Table 115</U></p></i></caption> |
<thead> | <thead> | ||
<tr> | <tr> | ||
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<h6><U>Results :</U></h6><br /><br /> | <h6><U>Results :</U></h6><br /><br /> | ||
<center><img src = "https://static.igem.org/mediawiki/2016/b/bb/Week_10_154Gel_extraction_of_A1-A2-D1-D2.jpg"; alt "Extraction gel of A1-A2-D1-D2"/> | <center><img src = "https://static.igem.org/mediawiki/2016/b/bb/Week_10_154Gel_extraction_of_A1-A2-D1-D2.jpg"; alt "Extraction gel of A1-A2-D1-D2"/> | ||
| − | <i><p> <U>Figure | + | <i><p> <U>Figure 19 :</U> Extraction gel of A1-A2-D1-D2 </p></i></center> |
<br /><br /> <br /> | <br /><br /> <br /> | ||
</p> | </p> | ||
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Follow the Qiagen Extraction gel kit steps with :<br /><br /> | Follow the Qiagen Extraction gel kit steps with :<br /><br /> | ||
<table> | <table> | ||
| − | <caption align="bottom" align="center">Table | + | <caption align="bottom" align="center"><i><p> <U>Table 116</U></p></i></caption> |
<thead> | <thead> | ||
<tr> | <tr> | ||
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1. Use 1/10 volume of NaAc and 2.5 colume of ethanol for each insert : <br /><br /> | 1. Use 1/10 volume of NaAc and 2.5 colume of ethanol for each insert : <br /><br /> | ||
<table> | <table> | ||
| − | <caption align="bottom" align="center">Table | + | <caption align="bottom" align="center"><i><p> <U>Table 117</U></p></i></caption> |
<thead> | <thead> | ||
<tr> | <tr> | ||
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Use the following volumes : <br /> | Use the following volumes : <br /> | ||
<table> | <table> | ||
| − | <caption align="bottom" align="center">Table | + | <caption align="bottom" align="center"><i><p> <U>Table 118</U></p></i></caption> |
<thead> | <thead> | ||
<tr> | <tr> | ||
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2. Digest the plasmid with the following volumes for each sample : <br /> | 2. Digest the plasmid with the following volumes for each sample : <br /> | ||
<table> | <table> | ||
| − | <caption align="bottom" align="center">Table | + | <caption align="bottom" align="center"><i><p> <U>Table 119</U></p></i></caption> |
<thead> | <thead> | ||
<tr> | <tr> | ||
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<h6><U>Results :</U></h6><br /><br /> | <h6><U>Results :</U></h6><br /><br /> | ||
<center><img src = "https://static.igem.org/mediawiki/2016/a/ad/Week_10_161Electrophoresis_of_the_PCR_done_on_the_8th_of_August_with_A1-A2-D1-D2.jpg" ; alt "Electrophoresis gel of the PCR"/> | <center><img src = "https://static.igem.org/mediawiki/2016/a/ad/Week_10_161Electrophoresis_of_the_PCR_done_on_the_8th_of_August_with_A1-A2-D1-D2.jpg" ; alt "Electrophoresis gel of the PCR"/> | ||
| − | <i><p> <U>Figure | + | <i><p> <U>Figure 20 :</U> Electrophoresis gel of the PCR</p></i></center><br /> |
The PCR works properly since we noticed significant bands at the expected level.<br /> | The PCR works properly since we noticed significant bands at the expected level.<br /> | ||
<br /><br /><br /> | <br /><br /><br /> | ||
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1. Realize a master mix with : <br /><br /> | 1. Realize a master mix with : <br /><br /> | ||
<table> | <table> | ||
| − | <caption align="bottom" align="center">Table | + | <caption align="bottom" align="center"><i><p> <U>Table 120</U></p></i></caption> |
<thead> | <thead> | ||
<tr> | <tr> | ||
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1. Put all the following reactants in a 1.5 ml Eppendorf and let digest one hour at 37°C : <br /> | 1. Put all the following reactants in a 1.5 ml Eppendorf and let digest one hour at 37°C : <br /> | ||
<table> | <table> | ||
| − | <caption align="bottom" align="center">Table | + | <caption align="bottom" align="center"><i><p> <U>Table 121</U></p></i></caption> |
<thead> | <thead> | ||
<tr> | <tr> | ||
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1. Make two measurements separated of 30 minutes : <br /> | 1. Make two measurements separated of 30 minutes : <br /> | ||
<table> | <table> | ||
| − | <caption align="bottom" align="center">Table | + | <caption align="bottom" align="center"><i><p> <U>Table 122</U></p></i></caption> |
<thead> | <thead> | ||
<tr> | <tr> | ||
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1. Start with tube 2 (8.6 ng/µl in 46 µl) and use the following mix : <br /> | 1. Start with tube 2 (8.6 ng/µl in 46 µl) and use the following mix : <br /> | ||
<table> | <table> | ||
| − | <caption align="bottom" align="center">Table | + | <caption align="bottom" align="center"><i><p> <U>Table 123</U></p></i></caption> |
<thead> | <thead> | ||
<tr> | <tr> | ||
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1. In a 1.5 ml Eppendorf, put : <br /> | 1. In a 1.5 ml Eppendorf, put : <br /> | ||
<table> | <table> | ||
| − | <caption align="bottom" align="center">Table | + | <caption align="bottom" align="center"><i><p> <U>Table 124</U></p></i></caption> |
<thead> | <thead> | ||
<tr> | <tr> | ||
