Team:Stanford/NotebookPage/9 July 2010
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Karpadillo (Talk | contribs) (→Karina's Notebook) |
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!!note sizes of plasmids/inserts <br/><br/> | !!note sizes of plasmids/inserts <br/><br/> | ||
- | GFP --> | + | GFP plasmid (pSB1A2) --> 2079 bp <br/> |
- | + | GFP insert --> 720 bp<br/> | |
+ | RFP plasmid (pSB2K3) --> 4425 bp <br/> | ||
+ | RFP insert --> 681 bp |
Revision as of 19:17, 19 July 2010
[http://docs.google.com/present/edit?id=0Ae31et9w_tAhZGZyc3R2ejJfOGNnY2drMmZz&hl=en&authkey=CPHGzfIB Weekly Leader Presentation: Karina]
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 plasmid (pSB1A2) --> 2079 bp
GFP insert --> 720 bp
RFP plasmid (pSB2K3) --> 4425 bp
RFP insert --> 681 bp