BioCurious: DIY Ultrasonic Transformation
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Revision as of 22:12, 18 March 2013
Experiment: Transform HB101 Strain with pGlo plasmid using ultrasound device
For an XX μL transformation:
Equipment: Off-the-shelf Ultrasonic Jewellery cleaning box with about 1L bath Strains DNA reagent in Dark Glass flat bottom tubes 5 mL with Plastic Caps
Sonicator Essentials: 42 KHz 35 watt peak 20 sq cm transducer area in center of bath
Bath filled with tap water at room temperature
- In a PCR tube, mix the components on ice in the order they are listed above.
- Perform thermocycling program
- 95 °C 5 min
- 95 °C 30 s
- TH 30 s
- 72 °C 1 min for each 1 kb PCR product
- Repeat steps 2-4 a total of 12-36 times (24 is standard).
- 72 °C 5 min
- 12 °C hold
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
- You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
- Anecdotal observations that might be of use to others can also be posted here.
Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
Relevant papers and books
- Goldbeter, A and Koshland, DE (1981) - Proc Natl Acad Sci U S A 78(11) 6840-4 PMID 6947258
- Jacob, F and Monod, J J (1961) - Mol Biol 3(3) 318-56 PMID 13718526
- Ptashne, M (2004) Genetic Switch: Phage Lambda Revisited - Cold Spring Harbor Laboratory Press ISBN 0879697164
- Who has experience with this protocol?
or instead, discuss this protocol.