Difference between revisions of "Diagnostic RP Digestion for Checking Insert Size"
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#Transfer about 300 ng into each tube and make up the difference with pure dH2O to 6 ul. Or just use 6 ul of a miniprep. | #Transfer about 300 ng into each tube and make up the difference with pure dH2O to 6 ul. Or just use 6 ul of a miniprep. | ||
#Prepare a master mix as follows: | #Prepare a master mix as follows: | ||
| − | + | 10 ng vector x (insert size/vector size) x 6 = ng insert, eg. 10ng x (700 bp/2100 bp) x 6 = 20 ng | |
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# | # | ||
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Revision as of 16:06, 21 June 2010
Diagnostic RP Digestion for Checking Insert Size
This procedure is for checking the insert size of a BioBrick part. It uses digestion with EcoRI + PstI.
- Label a 1.5 ml tube for each of the digestions to be set up.
- Transfer about 300 ng into each tube and make up the difference with pure dH2O to 6 ul. Or just use 6 ul of a miniprep.
- Prepare a master mix as follows:
10 ng vector x (insert size/vector size) x 6 = ng insert, eg. 10ng x (700 bp/2100 bp) x 6 = 20 ng
