BioCurious: DIY Ultrasonic Transformation

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Materials)
(Materials)
Line 7: Line 7:
For an XX μL transformation:
For an XX μL transformation:
-
Equipment:   
+
Equipment:  
-
Off-the-shelf Ultrasonic Jewellery cleaning box with about 1L bath
+
    
 +
Off-the-shelf Ultrasonic Jewellery cleaning box with ~ 1L bath
Strains  
Strains  
DNA
DNA
Line 15: Line 16:
Sonicator Essentials:  
Sonicator Essentials:  
 +
42 KHz
42 KHz
35 watt peak
35 watt peak

Revision as of 21:15, 18 March 2013

Contents

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. TH 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

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

  1. List troubleshooting tips here.
  2. You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
  3. 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.

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

  • Who has experience with this protocol?

or instead, discuss this protocol.

Personal tools