Difference between revisions of "Team:NTU-LIHPAO-Taiwan/Basic Part"

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	<div class="Text1" id="First5"><Red>DNA Ligation</Red></div>		<div class="Text1" id="First5"><Red>DNA Ligation</Red></div>
−	<div class="Text2">
−	<Green>Materials and Reagents :</Green>
−	</div>
−	<div class="Text3">
−	<ol class="part1">
−	<li>DNA (insert & vector)</li>
−	<li>Ligation high ver.2</li>
−	</ol>
−	</div>
−	<div class="Text2">
−	<Green>Equipment :</Green>
−	</div>
−	<div class="Text3">
−	<ol class="part1">
−	<li>Cooling dry bath incubator</li>
−	</ol>
−	</div>
	<div class="Text2">		<div class="Text2">
	<Green>Procedure :</Green>		<Green>Procedure :</Green>

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Revision as of 07:25, 7 September 2015

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• Notes
  • Preparation of Competent Cells (E.coli)
  • DNA Dissolution
  • Agarose Gel Electrophoresis
  • Gel Extraction
  • DNA Ligation
  • Plasmid DNA Extraction Using Alkaline Lysis Method (E.coli)
  • Transformation (E.coli)

DNA Ligation

Procedure :

1. Table

   Components	Volume (μL)
   Insert	x
   Vector	y
   Ligation High	1
   Total	7

           Molar ratio: insert/vector = 3/1
           x + y = 6
   
2. Gently mix the solution by pipetting up and down
3. Incubate
   1. at 16℃ for 2 hours, or
   2. at 37℃ for 1 hour, or
   3. at 4℃ overnight
4. Proceed with bacterial transformation

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