Stanford/BIOE44:Module 5:Day3: Difference between revisions
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==PCR amplifying your parts== | ==PCR amplifying your parts== | ||
Your primers have arrived! | |||
We might use this polymerase: [http://tools.invitrogen.com/content/sfs/manuals/pfx50_man.pdf Pfx50 DNA polyermase] It is higher fidelity than TAQ (which means it makes less mistakes). | |||
Revision as of 23:57, 22 May 2010
M5: Day 2 - Continue Building
Freezer Stocks of Synthesized Parts
Materials
- 5mL overnight culture
- sterile 60% glycerol
- 3 cryovials, labelled
Procedure
- Label your cryovials: part name, plasmid, date, your initials.
- Add 500ul of sterile 60% glycerol to each of your cryovials.
- Add 1.5mL of overnight culture to each cryovial. (Its easiest to use a 5ml pipette, take up 4.5mL of culture then drop 1.5mL into each tube)
- Invert tubes several times to mix.
- Put in freezer box in -80C freezer. (Usually one tube is kept as the working stock and the other two are kept as back ups in a separate freezer.)
PCR amplifying your parts
Your primers have arrived!
We might use this polymerase: Pfx50 DNA polyermase It is higher fidelity than TAQ (which means it makes less mistakes).
