BioCurious: DIY Ultrasonic Transformation

Overview

Experiment: Transform HB101 Strain with pGlo plasmid using ultrasound device

Materials

For an XX μL transformation:

Equipment:

Off-the-shelf Ultrasonic Jewellery cleaning box with ~ 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

Procedure

1. In a PCR tube, mix the components on ice in the order they are listed above.
2. Perform thermocycling program
   1. 95 °C 5 min
   2. 95 °C 30 s
   3. T_H 30 s
   4. 72 °C 1 min for each 1 kb PCR product
   5. Repeat steps 2-4 a total of 12-36 times (24 is standard).
   6. 72 °C 5 min
   7. 12 °C hold

---

Still in progress, above kept for template.

Sonicate each tube for 10 secs at only available setting on Sonicator (on/off)except 2nd control tube with plasmid and Al

1. Test transform Tube 1 with Al placed in middle of Transducer bump, flat glass bottom down
2. Test transform Tube 2 without Al placed in middle of Transducer bump same way
3. Test transform Tube 3 with Al placed in a side of the bath away from transducer
4. Test transform Tube 4 without Al placed in a side of the bath away from transducer
5. Control Tube 1 without plasmid and Al placed in middle of transducer bump
6. Control Tube 2 with plasmid and Al left alone

Post sonication

1. Add 1 mL LB solution
2. Incubate at 37C 1 hr on a bath-shaker
   1. Plate LB/AmpC after 1 hour and incubate for a day
   2. Check plates for colony in UV light next day

Notes

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References

Relevant papers and books

1. Goldbeter, A and Koshland, DE (1981) - Proc Natl Acad Sci U S A 78(11) 6840-4 PMID 6947258
2. Jacob, F and Monod, J J (1961) - Mol Biol 3(3) 318-56 PMID 13718526
3. Ptashne, M (2004) Genetic Switch: Phage Lambda Revisited - Cold Spring Harbor Laboratory Press ISBN 0879697164

Contact

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