SBB10Ntbk-DanielSedor: Difference between revisions
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[[Template:SBB-Protocols_PCR3 | SOEing PCR]]<br> | [[Template:SBB-Protocols_PCR3 | SOEing PCR]]<br> | ||
[[Template:SBB-Protocols_PCR2 | Cloning by PCR]]<br> | [[Template:SBB-Protocols_PCR2 | Cloning by PCR]]<br> | ||
[[Template:SBB-Protocols_Zymo3 | Zymo Gel Purification]]<br> | |||
[[Template:SBB-Protocols_Enz4 | Ligation of EcoRI/BamHI digests]] | [[Template:SBB-Protocols_Enz4 | Ligation of EcoRI/BamHI digests]] |
Revision as of 13:16, 17 February 2010
Construction of sbb10 PCR DS001/CA1600 on Bca1587 5-1 (420bp, gp=A) PCR CA1597/DS002 on Bca1587 5-6 (767bp, gp=B) ---------------------------- PCR DS001 and DS002 on A+B (1051 bp, EcoRI/BamHI) Sub into pBjk2741 (EcoRI/BamHI, 2170+910, L) Product is pBjk2741-sb10 {<SB100x!} ---------------------------- DS001 Forward oligo for cloning of <SB100x! CCATAgaattcATGagatctGGTAAATCTAAAGAAATCTCTCAGGAC CA1600 CGAAACGCAGACGAGCTTTTTTGTGACGGTTCTGGAGCAGCGGTTTTTT PCA assembly of sleepingBeauty (Bca1587) CA1597 ATGCTGGAAGAAACCGGTACCAAAGTTTCTATCTCTACCGTTAAACGTG PCA assembly of sleepingBeauty (Bca1587) DS002 Reverse oligo for cloning of <SB100x! ctgatGGATCCttaGTATTTGGTAGCGTTACCTT Part: gatctGGTAAATCTAAAGAAATCTCTCAGGACCTGCGTAAACGTATCGTTGACCTGCACAAATCTGGTTCTTCTCTGGGTGCTATCTCTAAACGTCTGGCTGTTCCGCGTTCTTCTGTTCAGACCATCGTTCGTAAATACAAACA CCACGGTACCACCCAGCCGTCTTACCGTTCTGGTCGTCGTCGTGTTCTGTCTCCGCGTGACGAACGTACCCTGGTTCGTAAAGTTCAGATCAACCCGCGTACCACCGCTAAAGACCTGGTTAAAATGCTGGAAGAAACCGGTACC AAAGTTTCTATCTCTACCGTTAAACGTGTTCTGTACCGTCACAACCTGAAAGGTCACTCTGCTCGTAAAAAACCGCTGCTCCAGAACCGTCACAAAAAAGCTCGTCTGCGTTTCGCTACCGCTCACGGTGACAAAGACCGTACCT TCTGGCGTAACGTTCTGTGGTCTGACGAAACCAAAATCGAACTGTTCGGTCACAACGACCACCGTTACGTTTGGCGTAAAAAAGGTGAAGCTTGCAAACCGAAAAACACCATCCCGACCGTTAAACACGGTGGTGGTTCTATCAT GCTGTGGGGTTGCTTCGCTGCTGGTGGTACCGGTGCTCTGCACAAAATCGACGGTATCATGGACGCTGTTCAGTACGTTGACATCCTGAAACAGCACCTGAAAACCTCTGTTCGTAAACTGAAACTGGGTCGTAAATGGGTTTTC CAGCACGACAACGACCCGAAACACACCTCTAAAGTTGTTGCTAAATGGCTGAAAGACAACAAAGTTAAAGTTCTGGAATGGCCGTCTCAGTCTCCGGACCTGAACCCGATCGAAAACCTGTGGGCTGAACTGAAAAAACGTGTTC GTGCTCGTCGTCCGACCAACCTGACCCAGCTGCACCAGCTGTGCCAGGAAGAATGGGCTAAAATCCACCCGACCTACTGCGAAAAACTGGTTGAAGGTTACCCGAAACGTCTGACCCAGGTTAAACAGTTCAAAGGTAACGCTAC CAAATACTAAG
SOEing PCR
Cloning by PCR
Zymo Gel Purification
Ligation of EcoRI/BamHI digests