Team:IIT Madras/Notebook/Protocols/Gel elusion

From 2011.igem.org

bar iGEM 2011 - Home Page Indian Institute of Technology - Madras

Gel Elusion


  • Prepare a 0.8% low melthing agarose gel.
  • Add 50 ul preparative reaction product into large wells.
  • Run the gel at 100 V for 30 – 45 mins.
  • Cut the gel with restricted DNA and keep it in an microfuge tube.
  • To the cut DNA, add 3 volumes of Chaotropic salt. Incubate at 50 C for 10 minutes.
  • Add 10 ul of GPS (Glass Powder Solution) for a PCR product and 15 ul for a vector and incubate at Room Temperature for 10 mins. Shake the tubes continuously so that GPS is in suspension and doesn’t settle down.
  • Centrifuge the tubes at 12000 rpm for 2 minutes and discard supernatant.
  • Wash pellet by adding 50 volumes of wash buffer. That is, 500 ul of Wash buffer for insert and 750 ul for vector.
  • Resuspend the pellet in 20 ul of water and heat at 50 C for 10 minutes. Centrifuge the tubes at 12000 rpm for 2 mins. Store supernatant at -20 C.