26/07/13

From 2013.igem.org

(Difference between revisions)
Line 28: Line 28:
*To calculate it we divided 200ng by the concentration of each sample  
*To calculate it we divided 200ng by the concentration of each sample  
*The rest of the volume was made up with water to give a total of 5ul.
*The rest of the volume was made up with water to give a total of 5ul.
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*The samples were them incubated in a water bath at 37C by an hour
+
*The samples were then incubated in a water bath at 37C by an hour
*Agorose Gel preparation
*Agorose Gel preparation
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**Boil for 3 minutes, 1 minute at a time.
**Boil for 3 minutes, 1 minute at a time.
**Let it cool and add 5ul of ethidium bromide.
**Let it cool and add 5ul of ethidium bromide.
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**Pour and let set for 30 mins.
+
**Pour and let it set for 30 mins.
-
**Adding dye and and marker
+
**Adding dye and marker
**Used orange g loading dye.
**Used orange g loading dye.
***2ul to each sample
***2ul to each sample

Revision as of 08:17, 31 July 2013

  1. purification of the limonene biobrick - BBa_24
  • Plasmid Volume
    • 1.1 - 40ul
    • 1.2 - 40ul
    • 1.3 - 46ul
    • 1.4 - 39ul
    • 1.5 - 41ul
    • 1.2 - 32ul
  • measured DNA concentration using nanodrop
    • DNA concentration (ng)
    • 1.1 - 131.8
    • 1.2 - 77.1
    • 1.3 - 56.8
    • 1.4 - 109
    • 1.5 - 129.1
    • 1.2 - 93.3
  • Restriction enzyme digest of the DNA samples
    • enzymes: XbaI, PstI
    • master mix:
      • Buffer - 14ul
      • H2O - 88.2ul
      • XbaI - 1.4ul
      • PstI - 1.4ul
      • Total volume - 105ul
  • Each sample contained 5ul of DNA and 15ul of the master mix.
  • The total amount of DNA in each sample was 200ng
  • To calculate it we divided 200ng by the concentration of each sample
  • The rest of the volume was made up with water to give a total of 5ul.
  • The samples were then incubated in a water bath at 37C by an hour
  • Agorose Gel preparation
      • 20ml of 5xTBE
      • 80ml of water
      • 0.8g of agarose
    • Boil for 3 minutes, 1 minute at a time.
    • Let it cool and add 5ul of ethidium bromide.
    • Pour and let it set for 30 mins.
    • Adding dye and marker
    • Used orange g loading dye.
      • 2ul to each sample
    • Used 1kb ladder, 5ul per 1 marker well.
  • Run the gel
  • Gel picture

IGEM lim restr dig 260713.jpg