Rensselaer/6 August 2008

Promoter Group

Buffer exchange/purification of XbaI/SpeI digests of mRFP and Fe promoter plasmids using Promega Wizard SV Gel and PCR Clean-Up System

Add 40 ul (mRFP digest)/60 ul (Fe promoter digest) of membrane binding solution. Wait 1'

Centrifuge max speed 1'. Discard flowthrough.

Add 700 ul membrane wash solution. Centrifuge max speed 1'. Discard flowthrough.

Add 500 ul membrane wash solution. Centrifuge max speed 5'. Discard flowthrough.

Centrifuge max speed 1'.

Transfer column to sterile 1.5 ml tube.

Add 50 ul nuclease free H2O to column. Wait 1'. Centrifuge max speed 1'.

[DNA] (using nanodrop spectrophotometer) mRFP=> 36 ng/ul Fe promoter=> 25 ng/ul