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From 2010.igem.org

Meeting Minutes

[http://docs.google.com/present/edit?id=0Ae31et9w_tAhZGZyc3R2ejJfOGNnY2drMmZz&hl=en&authkey=CPHGzfIB Weekly Leader Presentation: Karina]

Agenda

Karina's Notebook

Today's Goal: Digest GFP, RFP, and Terminators. Will do (GFP + T) and (RFP + T) ligations on monday.

Part #'s:
GFP: E0040 (cut at S and P)
RFP: E1010 (cut at S and P)
Term: B1006 (cut at X and P)

Recipe
12.0 uL DNA
1.0 uL each enzyme
5.0 uL NEB buffer 2
5 uL BSA (10x)
Sterile H20 to fill up to 50 uL

Mix and put 37º waterbath for 2 or more hours.

Making Gel
To make 50 mL of 0.8% agarose gel, add:
.4 g agarose
50 mL TAE (1x)
10 uL EtBr

Loading the Gel
add 10 uL loading dye (6x) to digests
add 40 uL of dye + digests to wells
don't forget to include wells with controls! (uncut plasmids)
load 10 uL ladder
run at 95 V for about an hour

• note sizes of plasmids/inserts
  
  

GFP --> pSB1A2 ( RFP --> pSB2K3