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Team:Stanford/NotebookPage/9 July 2010
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Karpadillo (Talk | contribs) (→Karina's Notebook) |
Karpadillo (Talk | contribs) (→Karina's Notebook) |
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50 mL TAE (1x)<br/> | 50 mL TAE (1x)<br/> | ||
10 uL EtBr<br/><br/> | 10 uL EtBr<br/><br/> | ||
| + | |||
| + | '''Loading the Gel''' <br/> | ||
| + | add 10 uL loading dye (6x) to digests <br/> | ||
| + | add 40 uL of dye + digests to wells <br/> | ||
| + | don't forget to include wells with controls! (uncut plasmids) <br/> | ||
| + | load 10 uL ladder <br/> | ||
| + | run at 95 V for about an hour <br/> | ||
| + | |||
| + | *note sizes of plasmids/inserts <br/><br/> | ||
| + | GFP --> pSB1A2 ( | ||
| + | RFP --> pSB2K3 | ||
Revision as of 19:06, 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 --> pSB1A2 ( RFP --> pSB2K3
