User:Matt Hartings/Notebook/AU Biomaterials Design Lab/2012/06/11: Difference between revisions
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==Description== | ==Description== | ||
# Add | # Four Samples for transformation | ||
## Novagen Test Plasmid into NovaBlue GigaSingles (recently purchased) | |||
## Novagen Test Plasmid into M15MA competent cells (recently made) - cells sent from David Tirrell at Caltech | |||
## A71M M8S M33S M103S Ascaris Hb into NovaBlue GigaSingles (recently purchased) | |||
## WT Ascaris Hb into NovaBlue GigaSingles (recently purchased) | |||
Plates: | |||
# 1.875 g LB | |||
# 1.5 g Agar | |||
# 75mL water | |||
# autoclave | |||
# 75uL of 100mg/mL ampicillin after mixture had cooled but not set | |||
Transformation protocol | |||
# Place plastic culture tubes on ice for 15 minutes | |||
# Place DNA for transformation on ice | |||
# After DNA has thawed, place cells on ice. (Note - want to minimize the time that the cells are out of freezer and on ice) | |||
# Add 1uL of DNA to the appropriately labeled culture tube and let sit in ice for 1 minute | |||
# Add cells (30uL, 40uL, or 50uL) to the tube on top of the DNA and gently pipette up and down to mix cells and DNA | |||
# Put tube back in ice for 4 minutes | |||
# Heat shock at 42C for 80seconds | |||
# Add 100uL of SOC media | |||
# Shake at 37C for 1 hour | |||
# Plate 50uL of culture media | |||
==Data== | ==Data== |
Revision as of 06:27, 12 June 2012
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ObjectiveTransform PCR and test plasmids into competent cells Description
Plates:
Transformation protocol
Data
NotesThis area is for any observations or conclusions that you would like to note.
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