User:Karmella Haynes/Notebook/BioBrick cloning/2013/02/23: Difference between revisions
From OpenWetWare
Line 8: | Line 8: | ||
==02/23/13== | ==02/23/13== | ||
<!-- Precede finished items with a checkmark ✓ --> | <!-- Precede finished items with a checkmark ✓ --> | ||
* Type IIS Assembly, PCR: LOV | * Type IIS Assembly, PCR: H2B-LOV+his | ||
---- | ---- | ||
'''Type IIS Assembly, PCR: LOV-H2B+his''' | '''Type IIS Assembly, PCR: LOV-H2B+his''' | ||
# H2B = 378 bp, | # 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 (18 bp) | |||
# pSB1A3 = ~2000, gg0001/ | # 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. | |||
PCR | PCR | ||
Line 27: | Line 28: | ||
| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| rowspan="7" | <u>Expected:</u><br>1. H2B = | | rowspan="7" | <u>Expected:</u><br>1. H2B = 378<br>2. LOV = 447<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] | ||
|- | |- |
Revision as of 11:57, 25 February 2013
Karmella's BioBrick Cloning | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
02/23/13
Type IIS Assembly, PCR: LOV-H2B+his
PCR
Golden Gate Reactions
Thermal cycling
|