There once was a sonic cleaner in the store for some 20€.I thought about buying it, but won't it damage the plasmids while transforming?????
There once was a sonic cleaner in the store for some 20€.I thought about buying it, but won't it damage the plasmids while transforming?????
Awesome!! How much ug plasmid did you use?
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Dude, very awesome!
Do you happen to know your plasmid concentration?
--A
Hi Guys,
Per Patrik's request...
Kyle Taylor,Jay Hanson and self performed the experiment here at Biocurious, initial results are encouraging..
Only deviations from paper were because of material at hand availibility and additional Al foil cutting in tube..
-shashank
Experiment: Transform HB101 Strain with pGlo plasmid using Ultrasound device
Equipment: Off the shelf Ultrasonic Jewellery cleaning box with about 1l bath.
Strains, DNA and reagent in Dark Glass flat bottom tubes 5ml with Plastic Caps
Sonicator Essentials:
42 KHz
35 watt Peak
20cm sq transducer area in center of bath
Bath filled with Tap water at room temperature
Transform Tubes:
Four Tranform tubes with
- 500ul of O.N. culture of HB101 strain
- 5ul of pGlo plasmid prepared thru miniprep
- 1ml of 75mM CaCl2
- Two tubes with Al foil discs (two none)
Two Control Tubes
- Same as above , but no plasmid, no Al
- Same as Transform, with plasmid and Al
Procedure:
Each Tube was sonicated for 10 secs at only available setting on Sonicator (on/off)
except 2nd control tube with plasmid and Al
- Test transform Tube 1 with Al placed in middle of Transducer bump, flat glass bottom down
- Test transform Tube 2 without Al placed in middle of Transducer bump same way
- Test transform Tube 3 with Al placed in a side of the bath away from transducer
- Test transform Tube 4 without Al placed in a side of the bath away from transducer
- Control Tube 1 without plasmid and Al placed in middle of transducer bump
- Control Tube 2 with plasmid and Al left alone
Post sonication
Add 1 ml LB solution
incubate at 37C 1 hr on a bath-shaker
Plate on LB and Amp agar after hour and incubate for a day
Check plates for colony in UV light next day
Results:
Test tube
- With HB/PGlo/Al/midTransducer: 9 Colonies
- with HB/PGlo/midtransducer: 4 colonies
- with HB /PGlo/Al/side placed: 3 colonies
- with HB/Pglo/side placed: 1 colony
Control
1. Just bacteria/midplaced: 0 colony
2. Bacteria/Pglo/Al no sonication: 0 colony
FWIW, we are pretty happy about it...open to all ideas to improve it.
To view this discussion on the web visit https://groups.google.com/d/msg/diybio/-/WGPjM3vH07QJ.