User:Karmella Haynes/Notebook/BioBrick cloning/2013/02/23: Difference between revisions
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'''Type IIS Assembly, PCR: LOV-H2B+his''' | '''Type IIS Assembly, PCR: LOV-H2B+his''' | ||
# H2B = 378 bp, 1A3/H2B fwd + H2B/LOV rev (Template: H2B-GFP from Addgene) | # H2B = 378 bp, 1A3/H2B fwd + H2B/LOV rev (Template: H2B-GFP from Addgene) | ||
# LOV = 429 bp, LOV fwd + LOV+his/1A3 rev (Template: LOV from Addgene); note: reverse primer adds a 6-his tag ( | # LOV = 429 bp, LOV fwd + LOV+his/1A3 rev (Template: LOV from Addgene); note: reverse primer adds a 6-his tag and TAA stop (21 bp) | ||
# pSB1A3 = ~2000, gg0001 v2 + gg0002 v2 | # pSB1A3 = ~2000, gg0001 v2 + gg0002 v2 | ||
* Be sure to use the H2B and LOV primers ordered on 2/4/2003. The names are redundant with a previous version. | * Be sure to use the H2B and LOV primers ordered on 2/4/2003. The names are redundant with a previous version. | ||
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| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| rowspan="7" | <u>Expected:</u><br>1. H2B = 378<br>2. LOV = | | rowspan="7" | <u>Expected:</u><br>1. H2B = 378<br>2. LOV = 450<br>3, 4. pSB1A3 = ~2000 | ||
| rowspan="7" | [[Image:KAH022313_gel1.jpg|200px|Hover name]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | | rowspan="7" | [[Image:KAH022313_gel1.jpg|200px|Hover name]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
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Revision as of 12:02, 25 February 2013
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02/23/13
Type IIS Assembly, PCR: LOV-H2B+his
PCR
Golden Gate Reactions
Thermal cycling
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