User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli

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==Project Description/Abstract==
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[[Image:engineering_pctfs.jpg|300px|thumb|right|]]
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'''Assemblies'''
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* Synthetic biology is an exciting new field that aims to create novel biological functions by artificially combining natural molecular parts. Rational design allows us to engineer the behaviors of protein-based drugs, in ways that we cannot accomplish with small molecule-based drugs. DNA assembly will be used to build a combinatorial library of over 200 synthetic transcription factors (Pc-Tfs) that will potentially activate therapeutic genes in cancer and stem cells. As time allows, a select group of Pc-TF protein will be purified and analyzed biochemically.
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# KAH213: (4) KAH54/(S/P)/430 + (1) KAH210/(X/P)/2398
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# KAH214: " + (2) KAH211/(X/P)/1321
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# KAH215: " + (3) KAH212/(X/P)/1621
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# KAH216: hPCD/(S/P)/186 ✓ + KAH210/(X/P)/2398
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# KAH217: " + KAH211/(X/P)/1321
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# KAH218: " + KAH212/(X/P)/1621
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# KAH207/V0200: KAH207/(N/S)/1836 ✓ + V0200/(N/S dp)/4442
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# KAH208/V0200: KAH208/(N/S)/1884 ✓ + "
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# KAH209/V0200: KAH209/(N/S)/1932 ✓ + "
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==Notes==
 
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* Brady Laughlin's Spring 2012 project in the Haynes Lab at Arizona State University
 
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* BME 394 Honors Research Project
 
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> Digests (Fermentas FD)<br>
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{| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table -->
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|-valign="top"
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| <u>Reagent</u> || <u>Volume</u> || &nbsp;
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| rowspan="9" | [[Image:KAH_071211_gel1.tif|200px|Cloning digests 7/12/11]]<br>30 μL/lane, 1% agarose -->
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|colspan="2" style="background-color: #F2F2F2;"|
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| DNA (plasmid) || up to 25 μL
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{{LnNotebookRecentChanges2}}
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|-
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| 10x buffer || 3.0
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| enzyme 1 || 1.0
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|-
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| enzyme 2 || 1.0
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| dH<sub>2</sub>O || ---
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| &nbsp; || 30 μL --> 37°C/ ~30 min.
|}
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__NOTOC__
__NOTOC__

Revision as of 21:02, 3 April 2012

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Assemblies

  1. KAH213: (4) KAH54/(S/P)/430 + (1) KAH210/(X/P)/2398
  2. KAH214: " + (2) KAH211/(X/P)/1321
  3. KAH215: " + (3) KAH212/(X/P)/1621
  4. KAH216: hPCD/(S/P)/186 ✓ + KAH210/(X/P)/2398
  5. KAH217: " + KAH211/(X/P)/1321
  6. KAH218: " + KAH212/(X/P)/1621
  7. KAH207/V0200: KAH207/(N/S)/1836 ✓ + V0200/(N/S dp)/4442
  8. KAH208/V0200: KAH208/(N/S)/1884 ✓ + "
  9. KAH209/V0200: KAH209/(N/S)/1932 ✓ + "


> Digests (Fermentas FD)

Reagent Volume   Cloning digests 7/12/11
30 μL/lane, 1% agarose -->
DNA (plasmid) up to 25 μL
10x buffer 3.0
enzyme 1 1.0
enzyme 2 1.0
dH2O ---
  30 μL --> 37°C/ ~30 min.


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