Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2013/02/08: Difference between revisions
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* Final assembly product: pSB1A3 left end / H2B / LOV +His tag/ pSB1A3 right end | * Final assembly product: pSB1A3 left end / H2B / LOV +His tag/ pSB1A3 right end | ||
* Primer design | * Primer design | ||
H2B | #H2B | ||
1A3/H2B fwd 5'-cacaccaCGTCTCa TAGA ATGCCAGAGCCAGCG | 1A3/H2B fwd 5'-cacaccaCGTCTCa TAGA ATGCCAGAGCCAGCG | ||
H2B/LOV rev 5'-cacaccaCGTCTCa CCAA CTTAGCGCTGGTGTA | H2B/LOV rev 5'-cacaccaCGTCTCa CCAA CTTAGCGCTGGTGTA | ||
LOV | #LOV | ||
LOV fwd 5'-cacaccaCGTCTCa TTGGCTACTACACTT | LOV fwd 5'-cacaccaCGTCTCa TTGGCTACTACACTT | ||
LOV+his/1A3 rev 5'-cacaccaCGTCTCa TAGT ttagtggtgatggtgatgatgAAGTTCTTTTGCCGC | LOV+his/1A3 rev 5'-cacaccaCGTCTCa TAGT ttagtggtgatggtgatgatgAAGTTCTTTTGCCGC | ||
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<center> | <center> | ||
[[Image: | [[Image:VN_Gel_2-8-13.png|Results of the digests are shown.]] | ||
[[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]] | [[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]] | ||
</center> | </center> | ||
<center> | <center> | ||
Results of the PCR product digests are shown | Results of the PCR product digests are shown. It was later found that the transparency used was blocking the UV light, so no bands could be seen. | ||
</center> | </center> | ||
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| PCR Product || 260|| 280 || 260/280 || ng/μL | | PCR Product || 260|| 280 || 260/280 || ng/μL | ||
|- | |- | ||
| H2B || | | H2B || 0.078|| 0.043|| 1.81 || 77.995 | ||
|- | |||
| LOV || 0.08 || 0.044 || 1.817 || 80.443 | |||
|- | |||
| pSB1A3 || 0.023 || 0.014 || 1.727 || 23.386 | |||
|- | |- | ||
|} | |} | ||
Revision as of 05:52, 18 April 2013
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February 8, 2013Type IIS Assembly - Making PCR Products
1A3/H2B fwd 5'-cacaccaCGTCTCa TAGA ATGCCAGAGCCAGCG H2B/LOV rev 5'-cacaccaCGTCTCa CCAA CTTAGCGCTGGTGTA
LOV fwd 5'-cacaccaCGTCTCa TTGGCTACTACACTT LOV+his/1A3 rev 5'-cacaccaCGTCTCa TAGT ttagtggtgatggtgatgatgAAGTTCTTTTGCCGC
PCR program:
Results of the PCR product digests are shown. It was later found that the transparency used was blocking the UV light, so no bands could be seen.
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