Melbourne/Lab BL Notebook
From 2007.igem.org
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==Aug 2007: Primer design for PCR stitching== | ==Aug 2007: Primer design for PCR stitching== | ||
In order to stitch Nt-SopII to ComP at the sites detailed [[Melbourne/Blue_Photosensor_Background#Chosen_Fusion_Sites|here]], the following primers were designed: | In order to stitch Nt-SopII to ComP at the sites detailed [[Melbourne/Blue_Photosensor_Background#Chosen_Fusion_Sites|here]], the following primers were designed: | ||
| + | -- Oligo 1 -- | ||
| + | Sequence Name: BL_Con1_s | ||
| + | Sequence: Cagtcggtgcggacatcgttgcttgtaagcaaagcttatac | ||
| + | -- Oligo 2 -- | ||
| + | Sequence Name: BL_Con1_as | ||
| + | Sequence: GTATAAGCTTTGCTTACAAGCAACGATGTCCGCACCGACTG | ||
| + | Number of Bases: 41 | ||
| + | -- Oligo 3 -- | ||
| + | Sequence Name: BL_Con2_s | ||
| + | Sequence: TcggtgcggacatcgctgCTTGTAAGCAAAGCTTATACCTTTG | ||
| + | Number of Bases: 43 | ||
| + | -- Oligo 4 -- | ||
| + | Sequence Name: BL_Con2_as | ||
| + | Sequence: CAAAGGTATAAGCTTTGCTTACAAGCAGCGATGTCCGCACCGA | ||
| + | Number of Bases: 43 | ||
| + | -- Oligo 5 -- | ||
| + | Sequence Name: BL_Con3_s | ||
| + | Sequence: GGTGCGGACATCGCTGGAGGTAAGCAAAGCTTATACCTTTGAG | ||
| + | Number of Bases: 43 | ||
| + | -- Oligo 6 -- | ||
| + | Sequence Name: BL_Con3_as | ||
| + | Sequence: CTCAAAGGTATAAGCTTTGCTTACCTCCAGCGATGTCCGCACC | ||
| + | Number of Bases: 43 | ||
| + | -- Oligo 7 -- | ||
| + | Sequence Name: BL_Con4_s | ||
| + | Sequence: GCGGACATCGCTGGAGGACAGCAAAGCTTATACCTTTGAGGT | ||
| + | Number of Bases: 42 | ||
| + | -- Oligo 8 -- | ||
| + | Sequence Name: BL_Con4_as | ||
| + | Sequence: ACCTCAAAGGTATAAGCTTTGCTGTCCTCCAGCGATGTCCGC | ||
| + | Number of Bases: 42 | ||
| + | -- Oligo 9 -- | ||
| + | Sequence Name: BL_Con5_s | ||
| + | Sequence: GACATCGCTGGAGGACGCCAAAGCTTATACCTTTGAGGTTACTC | ||
| + | Number of Bases: 44 | ||
| + | -- Oligo 10 -- | ||
| + | Sequence Name: BL_Con5_as | ||
| + | Sequence: GAGTAACCTCAAAGGTATAAGCTTTGGCGTCCTCCAGCGATGTC | ||
| + | Number of Bases: 44 | ||
| + | -- Oligo 11 -- | ||
| + | Sequence Name: BL_Con6_s | ||
| + | Sequence: gacgcagagcaggcacaaaagGTGATATTTTTAGATAAGCATGAAGTT | ||
| + | Number of Bases: 48 | ||
| + | -- Oligo 12 -- | ||
| + | Sequence Name: BL_Con6_as | ||
| + | Sequence: AACTTCATGCTTATCTAAAAATATCACCTTTTGTGCCTGCTCTGCGTC | ||
| + | Number of Bases: 48 | ||
| + | -- Oligo 13 -- | ||
| + | Sequence Name: BL_Con7_s | ||
| + | Sequence: GGCAGAGGAGATCAACACGGAAGTTGGACCGGACTGGAATT | ||
| + | Number of Bases: 41 | ||
| + | -- Oligo 14 -- | ||
| + | Sequence Name: BL_Con7_as | ||
| + | Sequence: AATTCCAGTCCGGTCCAACTTCCGTGTTGATCTCCTCTGCC | ||
| + | Number of Bases: 41 | ||
| + | -- Oligo 15 -- | ||
| + | Sequence Name: BL_FP1_s | ||
| + | Sequence: AGCTAGGaattcgcggccgcttctagATGGTGGGACTTACGACCCTCT | ||
| + | Number of Bases: 48 | ||
| + | -- Oligo 16 -- | ||
| + | Sequence Name: NL26-F | ||
| + | Sequence: GAATTCGCGGCCGCTTCTAGATAGAAGGATTTACGCGC | ||
| + | Number of Bases: 38 | ||
Revision as of 12:01, 10 October 2007
<Return to Lab notebook> <team home page>
Aug 2007: Order ComP and ComA from GENEART
We were counting on [http://partsregistry.org/Part:BBa_J51000 ComP] and [http://partsregistry.org/Part:BBa_J51001 ComA] being available in the 2007 iGEM distribution. As this was not the case, and we had DNA synthesis bases to burn, we ended up ordering them from GENEART. The original sequences, as described in the above links, were optimized for expression in e.coli and all cis-acting sites and assembly restriction sites were removed. We also included biobrick prefix/suffix and vf2 and vr primer binding sites. The two sequences, now labeled ComA' and ComP' took around 3 weeks after confirmation of order to be dispatched. The sequences have been uploaded below:
ComA' sequence (As ordered sequence)
ComP' sequence (As ordered sequence)
Aug 2007: Primer design for PCR stitching
In order to stitch Nt-SopII to ComP at the sites detailed here, the following primers were designed: -- Oligo 1 --
Sequence Name: BL_Con1_s Sequence: Cagtcggtgcggacatcgttgcttgtaagcaaagcttatac
-- Oligo 2 --
Sequence Name: BL_Con1_as Sequence: GTATAAGCTTTGCTTACAAGCAACGATGTCCGCACCGACTG Number of Bases: 41
-- Oligo 3 --
Sequence Name: BL_Con2_s Sequence: TcggtgcggacatcgctgCTTGTAAGCAAAGCTTATACCTTTG Number of Bases: 43
-- Oligo 4 --
Sequence Name: BL_Con2_as Sequence: CAAAGGTATAAGCTTTGCTTACAAGCAGCGATGTCCGCACCGA Number of Bases: 43
-- Oligo 5 --
Sequence Name: BL_Con3_s Sequence: GGTGCGGACATCGCTGGAGGTAAGCAAAGCTTATACCTTTGAG Number of Bases: 43
-- Oligo 6 --
Sequence Name: BL_Con3_as Sequence: CTCAAAGGTATAAGCTTTGCTTACCTCCAGCGATGTCCGCACC Number of Bases: 43
-- Oligo 7 --
Sequence Name: BL_Con4_s Sequence: GCGGACATCGCTGGAGGACAGCAAAGCTTATACCTTTGAGGT Number of Bases: 42
-- Oligo 8 --
Sequence Name: BL_Con4_as Sequence: ACCTCAAAGGTATAAGCTTTGCTGTCCTCCAGCGATGTCCGC Number of Bases: 42
-- Oligo 9 --
Sequence Name: BL_Con5_s Sequence: GACATCGCTGGAGGACGCCAAAGCTTATACCTTTGAGGTTACTC Number of Bases: 44
-- Oligo 10 --
Sequence Name: BL_Con5_as Sequence: GAGTAACCTCAAAGGTATAAGCTTTGGCGTCCTCCAGCGATGTC Number of Bases: 44
-- Oligo 11 --
Sequence Name: BL_Con6_s Sequence: gacgcagagcaggcacaaaagGTGATATTTTTAGATAAGCATGAAGTT Number of Bases: 48
-- Oligo 12 --
Sequence Name: BL_Con6_as Sequence: AACTTCATGCTTATCTAAAAATATCACCTTTTGTGCCTGCTCTGCGTC Number of Bases: 48
-- Oligo 13 --
Sequence Name: BL_Con7_s Sequence: GGCAGAGGAGATCAACACGGAAGTTGGACCGGACTGGAATT Number of Bases: 41
-- Oligo 14 --
Sequence Name: BL_Con7_as Sequence: AATTCCAGTCCGGTCCAACTTCCGTGTTGATCTCCTCTGCC Number of Bases: 41
-- Oligo 15 --
Sequence Name: BL_FP1_s Sequence: AGCTAGGaattcgcggccgcttctagATGGTGGGACTTACGACCCTCT Number of Bases: 48
-- Oligo 16 --
Sequence Name: NL26-F Sequence: GAATTCGCGGCCGCTTCTAGATAGAAGGATTTACGCGC Number of Bases: 38
