Sentelab:Notebook/gene synthesis/2010/09/18

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(Autocreate 2010/09/18 Entry for Sentelab:Notebook/gene_synthesis)
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==Entry title==
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==1st PCR product purification by PCR purification kit =
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* Insert content here...
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#5 volumes of buffer PB added to volume of the PCR reaction and mixed
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#A QIAquick column is plaed ina provided 2 ml collection tube
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#To bind DNA, the sample is applied to the QIAquick
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column and centrifuged for 45 sec. Flow trough is discarded and QIAquick column is placed back into the same tube.
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#To wash 0,75 ml buffer PE is added to the QIAquick column and centrifuged for 45 sec . Flow through is discaded and QIAquick column is placed back into the same tube.
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#The column is centrifuged in a 2ml collection tube for 1min
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#Each QIA quick column is placed in a clean 1.5 ml microcentrifuge tube.
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#The elute DNA for increased DNA concentration 30 ul EB buffer added to the center of the QIAquick membrane let the column stand for 1 min and than centrifuged.
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*REsults
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#1-11 - gfp oligos : 100,3 ng/uL
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#12-22 gfp oligos : 89,7 ng/uL

Revision as of 10:25, 17 September 2010

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=1st PCR product purification by PCR purification kit

  1. 5 volumes of buffer PB added to volume of the PCR reaction and mixed
  2. A QIAquick column is plaed ina provided 2 ml collection tube
  3. To bind DNA, the sample is applied to the QIAquick

column and centrifuged for 45 sec. Flow trough is discarded and QIAquick column is placed back into the same tube.

  1. To wash 0,75 ml buffer PE is added to the QIAquick column and centrifuged for 45 sec . Flow through is discaded and QIAquick column is placed back into the same tube.
  1. The column is centrifuged in a 2ml collection tube for 1min
  2. Each QIA quick column is placed in a clean 1.5 ml microcentrifuge tube.
  3. The elute DNA for increased DNA concentration 30 ul EB buffer added to the center of the QIAquick membrane let the column stand for 1 min and than centrifuged.
  • REsults
  1. 1-11 - gfp oligos : 100,3 ng/uL
  2. 12-22 gfp oligos : 89,7 ng/uL



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