Difference between revisions of "User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/01/24"

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(Autocreate 2013/01/24 Entry for User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli)
 
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=Date=
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=01/24/2013=
  
'''List title'''
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'Making Standardized DNA Part (NLS-HIS-STOP)'
# List items
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NLS-6His-STOP-F1 CTAGAcccaagaaaaagcgcaaggtacaccatcaccaccatcacgcgtaaagctgagACTAGTAGCGGCCGCTGCA 76
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NLS-6His-STOP-R1 GCGGCCGCTACTAGTctcagctttacgcgtgatggtggtgatggtgtaccttgcgctttttcttgggT 68
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Set up an annealing reaction as follows:
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{|
 +
|-
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| Sense oligo 1 (100 μM) || 3.0 μl
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|-
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| Anti-sense oligo  (100 μM) || 3.0 μl
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|-
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| 10x annealing buffer* || 2.0 μl
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|-
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| dH2O || 12.0 μl
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|-
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| &nbsp; || 20 μl
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|}
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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>
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--> *10x annealing buffer: 1 M NaCl; 100 mM Tris-HCl, pH 7.4

Revision as of 13:18, 24 January 2013

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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:

Sense oligo 1 (100 μM) 3.0 μl
Anti-sense oligo (100 μM) 3.0 μl
10x annealing buffer* 2.0 μl
dH2O 12.0 μl
  20 μl

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.
--> *10x annealing buffer: 1 M NaCl; 100 mM Tris-HCl, pH 7.4