User:Anugraha Raman/Notebook/iGEM 2010/2010/06/23: Difference between revisions

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**Scheme for PCR
**Scheme for PCR
[[Image: AmiRNA_pcr_scheme.jpg|thumb|center|amiRNA pcr diagram]]
[[Image: AmiRNA_pcr_scheme.jpg|thumb|center|amiRNA pcr diagram]]
**Go to the [[http://openwetware.org/wiki/IGEM:Harvard/2010/Team_Allergy| amiRNA section]] for our primers for this multi step PCR
**Go to the [[http://openwetware.org/wiki/IGEM:Harvard/2010/Team_Allergy| amiRNA section]] for our primers for this multi step PCR or see chart below:
 
{| class="wikitable" border ="1px solid black"
|-
|description
|forward (A)
|reverse (B)
|-
|biobrick RS300
|''CCTTGAATTCGCGGCCGCATCTAGA'' CCCACAAACACACGCTCGGA
|''AAGGCTGCAGCGGCCGCTACTAGT'' CCCCATGGCGATGCCTTAAA
|}
 
 
 
{| class="wikitable" border ="1px solid black"
|-
|description
|primer 1 (mir sense)
|primer 2 (mir antisense)
|primer 3 (mir* sense)
|primer 4 (mir* antisense)
|-
|[http://wmd3.weigelworld.org/cgi-bin/webapp.cgi?page=Designer&project=stdwmd&rm=show_results&id=7205 GFP]
|gaTTAGTGGTCGGCGAGCTGCACtctctcttttgtattcc
|gaGTGCAGCTCGCCGACCACTAAtcaaagagaatcaatga
|gaGTACAGCTCGCCGTCCACTATtcacaggtcgtgatatg
|gaATAGTGGACGGCGAGCTGTACtctacatatatattcct
|-
|[http://wmd3.weigelworld.org/cgi-bin/webapp.cgi?page=Designer&project=stdwmd&rm=show_results&id=7202 LTP 1]
|gaTCTAACTATGTATAGGACCACtctctcttttgtattcc
|gaGTGGTCCTATACATAGTTAGAtcaaagagaatcaatga
|gaGTAGTCCTATACAAAGTTAGTtcacaggtcgtgatatg
|gaACTAACTTTGTATAGGACTACtctacatatatattcct
|-
|[http://wmd3.weigelworld.org/cgi-bin/webapp.cgi?page=Designer&project=stdwmd&rm=show_results&id=7115 Bet v1]
|gaTTGGTTATATATATCGCACCTtctctcttttgtattcc
|gaAGGTGCGATATATATAACCAAtcaaagagaatcaatga
|gaAGATGCGATATATTTAACCATtcacaggtcgtgatatg
|gaATGGTTAAATATATCGCATCTtctacatatatattcct
|-
|}
 


'''Transformations'''
'''Transformations'''

Revision as of 11:51, 23 June 2010

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Wednesday June 23, 2010

amiRNA

amiRNA pcr diagram
    • Go to the [amiRNA section] for our primers for this multi step PCR or see chart below:
description forward (A) reverse (B)
biobrick RS300 CCTTGAATTCGCGGCCGCATCTAGA CCCACAAACACACGCTCGGA AAGGCTGCAGCGGCCGCTACTAGT CCCCATGGCGATGCCTTAAA


description primer 1 (mir sense) primer 2 (mir antisense) primer 3 (mir* sense) primer 4 (mir* antisense)
GFP gaTTAGTGGTCGGCGAGCTGCACtctctcttttgtattcc gaGTGCAGCTCGCCGACCACTAAtcaaagagaatcaatga gaGTACAGCTCGCCGTCCACTATtcacaggtcgtgatatg gaATAGTGGACGGCGAGCTGTACtctacatatatattcct
LTP 1 gaTCTAACTATGTATAGGACCACtctctcttttgtattcc gaGTGGTCCTATACATAGTTAGAtcaaagagaatcaatga gaGTAGTCCTATACAAAGTTAGTtcacaggtcgtgatatg gaACTAACTTTGTATAGGACTACtctacatatatattcct
Bet v1 gaTTGGTTATATATATCGCACCTtctctcttttgtattcc gaAGGTGCGATATATATAACCAAtcaaagagaatcaatga gaAGATGCGATATATTTAACCATtcacaggtcgtgatatg gaATGGTTAAATATATCGCATCTtctacatatatattcct


Transformations

  • Transformed ligations (V0210+LTP1) (V0210+Ger 3 sense) (V0210+Ger 3 antisense)w/turbo cells
  • Grew up colonies w/ RS300 for minipreps


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