SBB10Ntbk-DanielSedor: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Daniel Sedor (talk | contribs) |
Daniel Sedor (talk | contribs) |
||
Line 56: | Line 56: | ||
<br> | <br> | ||
===17 February 2010=== | ===17 February 2010=== | ||
2 PCR's according to the [[Template:SBB-Protocols_PCR3 | | 2 PCR's according to the [[Template:SBB-Protocols_PCR3 | Cloning by PCR]] protocol using oligos DS001/CA1600 (gp=A) and DS002/CA1597 (gp=B) from the sbb10 construction file. | ||
===22 February 2010=== | ===22 February 2010=== |
Revision as of 19:14, 22 February 2010
Construction Files
sbb10
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
sbb39
Eco/Bam transfer pBjh1601CK-Bjh1380 Sub into pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L) Product is pBjk2741-Bjh1380 {PmgtCB}
sbb40
Eco/Bam transfer pBjh1601CK-Bjh1858 Sub into pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L) Product is pBjk2741-Bjh1858 {<NIS!}
Lab Notes
17 February 2010
2 PCR's according to the Cloning by PCR protocol using oligos DS001/CA1600 (gp=A) and DS002/CA1597 (gp=B) from the sbb10 construction file.
22 February 2010
Performed Preparative Gel on PCR products sbb10 gp=A and gp=B. PCR products did not agree with predictions made in the construction file. Prepared two new PCR's.
24 February 2010
Filler