SBB10Ntbk-ZhenZongHuang: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
No edit summary |
||
Line 13: | Line 13: | ||
===sbb15=== | ===sbb15=== | ||
# [http://openwetware.org/wiki/Template:SBB-Protocols_PCR1 Wobble] | # [http://openwetware.org/wiki/Template:SBB-Protocols_PCR1 Wobble] | ||
# [http://openwetware.org/wiki/Template:SBB-Protocols_Zymo2 Small Zymol Clean up] | # [http://openwetware.org/wiki/Template:SBB-Protocols_Zymo2 Small Zymol Clean up] | ||
# [http://openwetware.org/wiki/Template:SBB-Protocols_Enz1 EcoR1/BamH1 Digest of Wobble Product] | # [http://openwetware.org/wiki/Template:SBB-Protocols_Enz1 EcoR1/BamH1 Digest of Wobble Product] |
Revision as of 15:40, 17 February 2010
Zhen Z. Huang 15:05, 16 February 2010 (EST)
Planning for Wed, Feb 17th, 2010
sbb31
- PCR
- Analytical gel
- Regular Zymol Clean up
- EcoR1/BamH1 Digest of PCR Product
- Zymol Gel Purification
- Ligation of EcoR1/BamH1 Digest
- Mapping
- Zymol Gel Purification
sbb15
- Wobble
- Small Zymol Clean up
- EcoR1/BamH1 Digest of Wobble Product
- Zymol Gel Purification
- Ligation of EcoR1/BamH1 Digest
- Mapping
- Zymol Gel Purification
Zhen Z. Huang 13:00, 18 February 2010 (EST)
Construction Files for my parts
sbb31
sbb31: CA42 origin of replication Source: pEC22-CA42 Target Sequence: agcacttcagcgcgccgtagcatcgataaacattacgggatggggcgaaactgccatctgttcgaaatgacgcgcaaatgggcttacagggcgattcgtcagggctggccagcattctcacagtggcttgatgccgtgattcagcgtgtcgaaatgtacaacgcatcgcttcccgttccgctttcacctcctgaatgtcgggctattggcaagagtattgcgaaatacacgcacaggaacttcacggcggaaactttcgcacagtatgtggctgatacgcacacgccagaaatacaggccaagagaggcaggaaaggtggcatcgctaaaggcgaagcctacgatgacaagcgtttcatggcgctatgtatgctggagaatggatattctcagaaagctattgcggcgatgttggaggtttctactcgaaccattcgaaactggaaaagcggaaaatagcctatatcagataacagcgcctttctggcgtttttttgagcagtaggtcttttgccg Vector: pBjk2741 Short description: oriCA42 Genbank reference: D30056.1 Family: Origin of Replication PCR ZZH001 and ZZH002 on pEC22-CA42 (547 bp, EcoRI/BamHI) Sub into pBjk2741-Bca1144 (EcoRI/BamHI, 2472+910, L) Product is pBca9523-sbb31 {oriCA42} ---------------------------- ZZH001 Forward oligo for cloning of oriCA42 ccataGAATTCatgAGATCTagcacttcagcgcgccgtag ZZh002 Reverse oligo for cloning of oriCA42 ctgatGGATCCcggcaaaagacctactgctc
sbb15
sbb15: phiC31 attB Source: Synthetic Target Sequence: tgacggtctcgaagccgcggtgcgggtgccagggcgtgcccttgggctccccgggcgcgtactccacctcacccatctggtcca Vector: pBjk2741 Short descriptions: phiC31 attB Genbank reference: AB306970 (759…842) Family: Phage att site good 20 bp gtgccagggcgtgcccttgg Wobble ZZH003/ZZH004 (115bp, EcoRI/BamHI) Sub into pBjk2741-Bca1144 (EcoRI/BamHI, 2170+910, L) Product is pBjk2741-sbb15 {phiC31 attB} ---- ZZH003 Forward construction of phiC31 attB basic part ccataGAATTCatgAGATCTtgacggtctcgaagccgcggtgcgggtgccagggcgtgcccttgg ZZH004 Reverse construction of phiC31 attB basic part ctgatGGATCCtggaccagatgggtgaggtggagtacgcgcccggggagcccaagggcacgccctggcac