SBB10Ntbk-ZhenZongHuang: Difference between revisions
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==[[User:Zhen Zong Huang|Zhen Z. Huang]] 15:05, 16 February 2010 (EST)== | |||
===Planning for Wed, Feb 17th, 2010=== | |||
===sbb31=== | |||
1. PCR | |||
2. Analytucal gel | |||
3. Zymol clean up | |||
4. digestion | |||
5. gel purification | |||
6. ligation | |||
7. mapping | |||
8. gel purification | |||
===sbb15=== | |||
1. Wobble | |||
2. Analytucal gel | |||
3. Zymol small clean up | |||
4. digestion | |||
5. gel purification | |||
6. ligation | |||
7. mapping | |||
8. gel purification | |||
==[[User:Zhen Zong Huang|Zhen Z. Huang]] 13:00, 18 February 2010 (EST)== | |||
==Construction Files for my parts== | ==Construction Files for my parts== | ||
===sbb31=== | ===sbb31=== | ||
Revision as of 23:11, 16 February 2010
Zhen Z. Huang 15:05, 16 February 2010 (EST)
Planning for Wed, Feb 17th, 2010
sbb31
1. PCR 2. Analytucal gel 3. Zymol clean up 4. digestion 5. gel purification 6. ligation 7. mapping 8. gel purification
sbb15
1. Wobble 2. Analytucal gel 3. Zymol small clean up 4. digestion 5. gel purification 6. ligation 7. mapping 8. 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 pBca9523-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
