User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/01/24: Difference between revisions
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'' | 'Making Standardized DNA Part (NLS-HIS-STOP)' | ||
NLS-6His-STOP-F1 CTAGAcccaagaaaaagcgcaaggtacaccatcaccaccatcacgcgtaaagctgagACTAGTAGCGGCCGCTGCA 76 | |||
NLS-6His-STOP-R1 GCGGCCGCTACTAGTctcagctttacgcgtgatggtggtgatggtgtaccttgcgctttttcttgggT 68 | |||
Set up an annealing reaction as follows: | |||
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| Sense oligo 1 (100 μM) || 3.0 μl | |||
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| Anti-sense oligo (100 μM) || 3.0 μl | |||
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| 10x annealing buffer* || 2.0 μl | |||
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| dH2O || 12.0 μl | |||
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| || 20 μl | |||
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Heat at 100°C for 5 min., remove the entire heat block or water bath from the heat source, and allow to cool slowly to room temperature.<br> | |||
--> *10x annealing buffer: 1 M NaCl; 100 mM Tris-HCl, pH 7.4 | |||
Revision as of 14:18, 24 January 2013
 PcTF Subcloning in E. coli
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01/24/2013'Making Standardized DNA Part (NLS-HIS-STOP)' NLS-6His-STOP-F1 CTAGAcccaagaaaaagcgcaaggtacaccatcaccaccatcacgcgtaaagctgagACTAGTAGCGGCCGCTGCA 76 NLS-6His-STOP-R1 GCGGCCGCTACTAGTctcagctttacgcgtgatggtggtgatggtgtaccttgcgctttttcttgggT 68 Set up an annealing reaction as follows:
Heat at 100°C for 5 min., remove the entire heat block or water bath from the heat source, and allow to cool slowly to room temperature. |
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