Team:Toronto/Experiment-iGEM DNA Kit Plate Instructions

iGEM DNA Kit Plate Instructions · Benchling

iGEM DNA Kit Plate Instructions

Introduction

This protocol is taken directly from: http://parts.igem.org/Help:2016_DNA_Distribution#DNA_Distribution_Kit_Plates

Materials

  • iGEM DNA Kit
    • dH2O

      Procedure

      • To use the DNA in the Distribution Kit, follow these instructions:
      • Note: There is an estimated 2-3ng of DNA in each well, following this protocol, assume that you are transforming with 200-300pg/ul
      1. With a pipette tip, punch a hole through the foil cover into the corresponding well of the part that you want. Make sure you have properly oriented the plate. Do not remove the foil cover, as it could lead to cross contamination between the wells.
      1. Pipette 10uL of dH2O (distilled water) into the well. Pipette up and down a few times and let sit for 5 minutes to make sure the dried DNA is fully resuspended. The resuspension will be red, as the dried DNA has cresol red dye. We recommend that you do not use TE to resuspend the dried DNA.
      1. Transform 1ul of the resuspended DNA into your desired competent cells, plate your transformation with the appropriate antibiotic* and grow overnight.
      1. Pick a single colony and inoculate broth (again, with the correct antibiotic) and grow for 16 hours.
      1. Use the resulting culture to miniprep the DNA AND make your own glycerol stock (for further instruction on making a glycerol see this page). We recommend using the miniprepped DNA to run QC tests, such as restriction digests and sequencing.



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