Construction

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<font size=4>< [[Aug11-Sep7]] | [[Sep8-14]] | [[Sep15-26]] | [[Sep27-Oct3]] | [[Construction]] ></font>
<font size=4>< [[Aug11-Sep7]] | [[Sep8-14]] | [[Sep15-26]] | [[Sep27-Oct3]] | [[Construction]] ></font>
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</center>
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== October 11, 2006 ==
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'''To-Do List:'''
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<ul>
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    <li>Digest I12006 H (2005) and I12006 AB (2006) with XbaI/SpeI
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    <li>Transform cells with I12006 AB (2005)
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    <li>Transform cells with UT1 AB, UT2 EF, UT4 AB
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        <ul>
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            <li>Try to find the UT4 used to make a measurement (that was successful)
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        </ul>
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    <li>Digest I12006 AB (2005) with EcoRI/XbaI (host) and UT4 AB with EcoRI/SpeI (insert)
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        <ul>
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            <li>Ligate I12006 and UT4
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        </ul>
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    <li>Need to test to see if arabinose is having deleterious effects on fluorescence
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        <ul>
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            <li>DH5a auto-fluorescence vs. [Arabinose]
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            <li>Consider using a tetR promoter instead of pBad/araC
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        </ul>
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    <li>Once the mini-preps of the correct lengths are obtained, retest UT2/UT3 with increasing IPTG to repeat previous experiment (if
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        successful, take pictures)
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</ul>
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[http://2006.igem.org/University_of_Toronto_2006 Home]
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== October 8, 2006 ==
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'''Anne:'''
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<ul>
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    <li>Prepared DH5a UT2 for LacI temperature test
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    <li>Mini-prepped I2006 H (2005) and I12006 AB (2006)
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</ul>
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[http://2006.igem.org/University_of_Toronto_2006 Home]
== October 7, 2006 ==
== October 7, 2006 ==

Revision as of 23:56, 11 October 2006

< Aug11-Sep7 | Sep8-14 | Sep15-26 | Sep27-Oct3 | Construction >

Contents

October 11, 2006

To-Do List:

  • Digest I12006 H (2005) and I12006 AB (2006) with XbaI/SpeI
  • Transform cells with I12006 AB (2005)
  • Transform cells with UT1 AB, UT2 EF, UT4 AB
    • Try to find the UT4 used to make a measurement (that was successful)
  • Digest I12006 AB (2005) with EcoRI/XbaI (host) and UT4 AB with EcoRI/SpeI (insert)
    • Ligate I12006 and UT4
  • Need to test to see if arabinose is having deleterious effects on fluorescence
    • DH5a auto-fluorescence vs. [Arabinose]
    • Consider using a tetR promoter instead of pBad/araC
  • Once the mini-preps of the correct lengths are obtained, retest UT2/UT3 with increasing IPTG to repeat previous experiment (if successful, take pictures)

Home

October 8, 2006

Anne:

  • Prepared DH5a UT2 for LacI temperature test
  • Mini-prepped I2006 H (2005) and I12006 AB (2006)

Home

October 7, 2006

Cheng:

  • Prepared DH5a-z1 UT2 for IPTG induction of GFP

Conrad:

  • Re-test UT4C, UT5 ABC and I12006 EFG (2005) with XbaI/SpeI
  • Miniprep UT2/3 in DH5a

Siva:

  • Prepared DH5a UT2 for Arabinose induced LacI test

Recorded digest length check results:

  • I12006 EFG (4000-5000, 1500-1000)
  • UT5 ABC (3500-4000, 750-1000)
  • UT4 C (3000-3500)
  • UT3 DH5a (750-1000, 2000-2500)
  • UT2 DH5a (750-1000, 2000-2500)

Test Results

Home

October 6, 2006

Andy, HoKwon:

  • Prepared o/n (3 vials each) of DH5a and DH5a-z1 with UT2 and UT3 for testing
  • Prepared o/n of UT2 DH5a and UT3 DH5a for miniprep
  • Transformed and plated I12006 (2005) (poured in amp plate first by accident), I12006 (2006)
  • Double Digest parts E0240 CD (2005), UT4 AB with XbaI/SpeI
    • E0240 CD (2000-2500, 750-1000) Correct!
    • UT4 AB (3000-4000) Correct!

Charles, Cheng, Nick:

  • Re-tested UT4 C, UT5 ABC – failed due to a very dry gel

Home

October 4, 2006

Andy:

  • Checked lengths (EcoRI, PstI) of following
    • I12006 EFG (4000-5000, 1000-1500 for all three)
    • UT4 AB (3000-4000, 2000) C (3000-4000, 250)
    • UT5 ABC (2500-3000, 750-1000)
  • Digested following
    • I12006 (2005) E (6000 by SpeI and PstI)
    • E0240 (2005) C (2500-3000, 2000-2500, 1500-2000, 750-1000 by XbaI and PstI)
    • UT4 AB (6000-10000, 2000-2500 by SpeI and PstI) C (5000-6000, 3000 by SpeI and PstI)
    • UT5 ABC (3000-4000 by SpeI and PstI)
  • Gel extracted E0240 (insert) and UT4 AB (host)
  • Transformed and plated DH5a with UT2/UT3 for testing
  • Made o/n of UT2/UT3 with IPTG

To-Do:

  • Redo transformation of I12006 (2005)
  • Learn to take fluorescent images under microscope
  • Temperature testing
  • Try ligating UT5 again
  • Running out of water and PCR tubes

Home

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