Supplemental
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Adam.Emrich (Talk | contribs) (→Ligation) |
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</style> | </style> | ||
<body> | <body> | ||
- | <table | + | <table cellpadding=0 cellspacing=0 border=0> |
<tr> | <tr> | ||
<td width=682 align=left valign=top> | <td width=682 align=left valign=top> | ||
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===Restriction Digest=== | ===Restriction Digest=== | ||
- | 1. Combine the following ingredients: | + | <html>1. Combine the following ingredients: |
+ | |||
a. 1 uL of each restriction enzyme to be used. | a. 1 uL of each restriction enzyme to be used. | ||
b. 500-5000 ng of DNA. | b. 500-5000 ng of DNA. | ||
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f. 5 uL of Antarctic Phosphatase Buffer. | f. 5 uL of Antarctic Phosphatase Buffer. | ||
g. Enough H2O for 50 uL overall volume. | g. Enough H2O for 50 uL overall volume. | ||
+ | </html> | ||
2. Incubate at 37 C for 2-4 hours (or overnight). | 2. Incubate at 37 C for 2-4 hours (or overnight). | ||
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There are two options for ligations, 3A and standard. These are more general protocols, as we did not find a specific way to do them with a high rate of success. | There are two options for ligations, 3A and standard. These are more general protocols, as we did not find a specific way to do them with a high rate of success. | ||
- | + | 3A Ligations | |
1. For these ligations the following are necessary: | 1. For these ligations the following are necessary: | ||
a. The prefix part cut with EcoRI and SpeI. | a. The prefix part cut with EcoRI and SpeI. | ||
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4. Plate out the cells on a plate with an antibiotic corresponding to resistance of the construction plasmid. | 4. Plate out the cells on a plate with an antibiotic corresponding to resistance of the construction plasmid. | ||
- | + | Standard Ligations | |
1. After Ethanol Precipitation of Digested parts, run DNA through gel to extract parts of correct length. | 1. After Ethanol Precipitation of Digested parts, run DNA through gel to extract parts of correct length. | ||
2. Ethanol Precipitate these parts. | 2. Ethanol Precipitate these parts. | ||
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8. Use them to Transform competent cells. | 8. Use them to Transform competent cells. | ||
9. Plate cells with appropriate antibiotics. | 9. Plate cells with appropriate antibiotics. | ||
+ | </html> | ||
== '''Presentations''' == | == '''Presentations''' == | ||
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[[Media:Brown_iGEM_6-15-07.ppt]] | [[Media:Brown_iGEM_6-15-07.ppt]] | ||
- | ==Videos== | + | =='''Videos'''== |
[[Image:BrowniGEMYoutube2.jpg|thumb|left|4 minute Intro to iGEM http://youtube.com/watch?v=XpAhZd0Xu40]] [[Image:BrowniGEMYoutube1.jpg|thumb|left|Nanodrop it like it's hot! http://youtube.com/watch?v=pKT4Zrjeh6s]] | [[Image:BrowniGEMYoutube2.jpg|thumb|left|4 minute Intro to iGEM http://youtube.com/watch?v=XpAhZd0Xu40]] [[Image:BrowniGEMYoutube1.jpg|thumb|left|Nanodrop it like it's hot! http://youtube.com/watch?v=pKT4Zrjeh6s]] |
Latest revision as of 04:12, 27 October 2007
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