Haynes Lab:Notebook/Engineering PC-TFs/2013/10/24: Difference between revisions
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|style="background-color: #800000" align="center"| | |style="background-color: #800000" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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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. | |||
{| {{table}} | |||
| align="center" style="background:#efefef;"| '''Part''' | |||
| align="center" style="background:#efefef;"|'''Concentration''' | |||
|- | |||
| KAH126||143.967 ng/μL | |||
|} | |||
<big>''Cell Culture of KAH126''</big> | |||
'''Grow liquid culture of KAH126''' | |||
*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. | |||
*Grow the cultures for 10-12 hrs. | |||
Latest revision as of 23:30, 26 September 2017
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October 24, 2013Sigma-Aldrich Mini-prep Kit
Cell Culture of KAH126 Grow liquid culture of KAH126
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