Talk:Team:IvyTech-South Bend/9 September 2010

From 2010.igem.org

9/9/10

Due to conflict we will be changing from E Coli to yeast. Today we will electroporate the 2 parts we discovered yesterday parts #’s BBa_F2620 a pops receiver and # BBa_T9002 a receiver device. Both parts are on plate 2 of the Igems registry F2620 is well 6 – E, and T9002 is in well 9 – A Both parts.

•We will need to follow protocol from openwetware.org Refer to IGems Main Lab notebook for std. protocol.

•The DNA will be extracted inside Bio-fume hood • Before extraction I cleaned entire Fume hood with ethanol alcohol to sterilize * • Then placed sterile 1 mL centrifuge tube container into biohood • Set up electroporation machine set to EC - 1 setting and waiting • I took and added 10 mL 180 ohm H2O to well A – 9 • then pipetted up and down using my BR – 10 1 0 0 then pipetted 2 mL Ligated DNA into 100 mL electrocomp cells • added 100 mL to chilled on ice bath cuvette then electroporated hitting then button twice at 1.8 kv and into cuvette after electroporation 900 mL 50B media placed into cuvette then drew all 1 mL out and placed into 1.5 mL centrifuge tube. • 4.6 ms at 2:50 pm they were placed into a 1.5 mL centrifuge tube to stand for 1 hr. • After standing for 1 hr we will plate 100 mL transformed electro cells on LB/Agar with 100 mg/mL Amp to grow overnight at 30 degrees C