User:Karmella Haynes/Notebook/BioBrick cloning/2015/04/06: Difference between revisions
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Part 2: Prep the DNA fragments | Part 2: Prep the DNA fragments (PCR) | ||
* Note: Final conc. in LCR rxn is 3 nM each | * Note: Final conc. in LCR rxn is 3 nM each | ||
* Use '''primers with 5' phosphates''' to amplify the fragment(s) of interest in 50 μL reactions. Error-free PCR is highly recommended for large fragments. | * Use '''primers with 5' phosphates''' to amplify the fragment(s) of interest in 50 μL reactions. Error-free PCR is highly recommended for large fragments. | ||
* Digest the template DNA: Add 1 μL FastDigest DpnI and 5μL of 10x FastDigest buffer to each PCR reaction. Incubate at 37°C for 15 min. | * Digest the template DNA: Add 1 μL FastDigest DpnI and 5μL of 10x FastDigest buffer to each PCR reaction. Incubate at 37°C for 15 min. |
Revision as of 15:22, 3 April 2015
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04/06/15
LCR Continued
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