Team:Newcastle/Minipreps

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(New page: ==Minipreps== ===Gel extraction=== #Excise the DNA fragment from the agarose gel with a clean, sharp scalpel #Weigh the gel slice in the colourless tube. Add 3 volumes Buffer QG to 1 vol...)
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==Minipreps==
==Minipreps==

Revision as of 13:37, 28 July 2010

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Minipreps

Gel extraction

  1. Excise the DNA fragment from the agarose gel with a clean, sharp scalpel
  2. Weigh the gel slice in the colourless tube. Add 3 volumes Buffer QG to 1 volume of gel slice in a colourless tube (100mg~10µl)
  3. Incubate at 50°C for 10 min (or until the gel slice has completely dissolved)vortex every 2-3minutes
  4. After the gel slice has dissolved completely, check the colour of the mixture is yellow
  5. Add 1gell volume of isopropanol to the sample and mix
  6. Place QG quick spin column in 2ml collection tube
  7. Add sample and spin for 1 minute (discard flow through)
  8. Add QG buffer and spin for 1min (discard flow through)
  9. Add PE buffer and spin for 1min (discard flow through)(13000rpm)
  10. Place spin column in clean tube(1.5ml)
  11. Add elution buffer (50µl)spin for 1 min
  12. If using a gel to analyse add loading dye and mix (pipet up and down)