Haynes Lab:Notebook/Engineering PC-TFs/2013/10/24: Difference between revisions
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# <big>''Elute DNA''</big>: '''Transfer the column to a fresh collection tube. Add 100 µl of Elution Solution or molecular biology reagent water to the column.''' For DNA sequencing and other enzymatic applications, use water or 5 mM Tris-HCl, pH 8.0, as an eluant. '''Centrifuge at 12,000 x g for 1 minute.''' The DNA is now present in the eluate and is ready for immediate use or storage at –20 °C. Note: If a more concentrated plasmid DNA preparation is required, the elution volume may be reduced to a minimum of 50 µl. | # <big>''Elute DNA''</big>: '''Transfer the column to a fresh collection tube. Add 100 µl of Elution Solution or molecular biology reagent water to the column.''' For DNA sequencing and other enzymatic applications, use water or 5 mM Tris-HCl, pH 8.0, as an eluant. '''Centrifuge at 12,000 x g for 1 minute.''' The DNA is now present in the eluate and is ready for immediate use or storage at –20 °C. Note: If a more concentrated plasmid DNA preparation is required, the elution volume may be reduced to a minimum of 50 µl. | ||
<big>''Measure concentration of each fragment''</big>: | |||
* Load Gen 5 2.0 software. Go to the Task Manager-Take 3 Application | |||
* Go to Nucleic Acid Quantification. | |||
* Pipette 2 µL of each DNA sequence into Take 3 plate. | |||
* Place plate in EPOCH plate reader. | |||
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<big>''Cell Culture of KAH126''</big> | |||
''Grow liquid culture of KAH126'' | '''Grow liquid culture of KAH126''' | ||
*Label 15 ml sterile culture tubes. Fill each tube with <big>'''6 ml'''</big> LB growth medium with Amp. | *Label 15 ml sterile culture tubes. Fill each tube with <big>'''6 ml'''</big> LB growth medium with Amp. | ||
*Use sterile pipette tip to touch bacterial streak and put the tip into LB medium. | *Use sterile pipette tip to touch bacterial streak and put the tip into LB medium. |
Revision as of 14:06, 25 October 2013
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October 24, 2013Sigma-Aldrich Mini-prep Kit
Cell Culture of KAH126 Grow liquid culture of KAH126
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