User:Karmella Haynes/Notebook/BioBrick cloning/2013/03/14: Difference between revisions
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'''New mammalian expression vector MV9 | '''New mammalian expression vector MV9'''<br> | ||
* Use the [http://gcat.davidson.edu/igem10/ Oligator tool] to design oligo-assembly inserts for "MV6" (pcDNA3.1+ puro, mammalian transfection vector; CMV promoter) | * Use the [http://gcat.davidson.edu/igem10/ Oligator tool] to design oligo-assembly inserts for "MV6" (pcDNA3.1+ puro, mammalian transfection vector; CMV promoter) | ||
* MV9 - will carry Kozak-XbaI-NLS-6His-stop (for Brady's and Behzad's projects) | * MV9 - will carry Kozak-XbaI-NLS-6His-stop (for Brady's and Behzad's projects) | ||
** The XbaI site will be used to accept X/S inserts; plasmid digests will be needed to check proper orientation | |||
* Oligo assembly method will be used to make double stranded DNA with SpeI overhangs | * Oligo assembly method will be used to make double stranded DNA with SpeI overhangs | ||
* Insert will be ligated to XbaI-cut MV6 to destroy the preexisting XbaI site | * Insert will be ligated to XbaI-cut MV6 to destroy the preexisting XbaI site | ||
* Clones will be screened for proper orientation and insert copy number | * Clones will be screened for proper orientation and insert copy number | ||
Oligator results: | Oligator results: | ||
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<font face="courier"> | <font face="courier"> | ||
<u> | <u>ctagtCCCGCCGCCACCATGGAGTCTAGAC</u>CCAAGAAAAAGCGCAAGGTACACCATCACCACCATCACGCGTAAAGCTGAGa<br> | ||
<u> | aGGGCGGCGGTGGTACCTCAGATCTGGGTTCTTTTTCGCGTTCCATGTGGTA<u>GTGGTGGTAGTGCGCATTTCGACTCtgatc</u></font><br> | ||
# 30-mer 5'-ctagtCCCGCCGCCACCATGGAGTCTAGAC | # 30-mer 5'-ctagtCCCGCCGCCACCATGGAGTCTAGAC |
Revision as of 13:45, 9 April 2013
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04/14/13
New mammalian expression vector MV9
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