Upgrading old Caralina Electrophoresus for Dye

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bigmack
Posts: 816
Joined: 15 Dec 2015, 10:49
Job Title: Lab Technician
School: FCAC
State/Location: QLD

Upgrading old Caralina Electrophoresus for Dye

Post by bigmack »

We have had an older DNA electrophoresis kit .It originally cost around $500 . Then we bought a replacement kit at just under $200 .( this contains Another lot of Cut DNA samples , Carbon fibre electrodes , TBA buffer sol"n , stain and the ridiculously expensive Agarose powder .
Twice in the last two years it's been dragged out and teacher ended up with poor or no results .
I had the job of retesting it all and we concluded that the DNA samples must be completely useless due to their age .
Carolina Electrophoresus kit.jpg
The Head teacher said "no way can we spend $200 for a fresh lot of samples and expendables every time a teacher wants to do this Prac".
So We decided to get the Dyes from Southern Biological and give that a go as it only requires an electrolyte of Sodium Bicarbonate instead of the TBA buffer Solution . Also standard agar powder can be used instead of the Agarose .

Dye Electrophoresis is slightly different to DNA Electrophoresis in that the Dyes can have either a positive or negative charge so needs to be positioned in the middle of the Gel unlike DNA which is started at one end .
Alas out trays only had slots at the end to fit the well combs so I had to carefully make slots in the middle to suit the new comb position .This involved cutting a slot with a jeweler's saw to equal length as the original so that the combs set into the Gel the same depth
Setting agar.jpg
Another issue with our old trays is that they require a carbon fibre electrode which has to be cut to fit each use and then thrown away.
(Now all these problems could have been avoided if we just shelled out $185 each for new trays with Platinum electrodes built in . )...but that wasn't going to happen . [-X
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Last edited by bigmack on 17 May 2016, 19:02, edited 1 time in total.
bigmack
Posts: 816
Joined: 15 Dec 2015, 10:49
Job Title: Lab Technician
School: FCAC
State/Location: QLD

Re: Upgrading old Caralina Electrophoresus for Dye

Post by bigmack »

original carbon fibre electrodes.jpg
Anyways, I made up some electrodes out of carbon electrodes we had here and secured them in place with a copper clip which the alligator leads clip to .
carbon electrodes.jpg
Testing carbon electrodes.jpg
This means we no longer need to buy the replacement kits @ $200 to run this prac and can now do it with a $27 set of Dyes that will give us around 45 tests per dye sample....with 5 Gel trays , that's about 9 pracs :D

The only other thing I'm looking at is making a voltage multiplier circuit to fit to our Power supplies so we can get the required 40 odd volts to run this . In previous years we had multiple 9 volt batteries in series ....another huge cost ..

Or we could just buy the correct power supply , its only around $400 :cheesy:
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Merilyn1
Posts: 1476
Joined: 12 Mar 2013, 08:10
Job Title: Labbie
School: Wollondilly Anglican College
Suburb: Tahmoor
State/Location: NSW

Re: Upgrading old Caralina Electrophoresus for Dye

Post by Merilyn1 »

Wow, well done! You can hang out in my prep room any time you like - I'll give you all my "problem" bits of equipment to sort out.
bigmack
Posts: 816
Joined: 15 Dec 2015, 10:49
Job Title: Lab Technician
School: FCAC
State/Location: QLD

Re: Upgrading old Caralina Electrophoresus for Dye

Post by bigmack »

Thanks for the offer Merilyn :party:
LOL , but seriously it was Athletics Carnival yesterday and there is only sooo much Labeling you can do :whistling2:

I hate seeing expensive gear sitting around either because its not working or it's too expensive to run. The Dept had already thrown a good $750-$800 at this kit for effectively two successful Pracs .
If I had not done what I did it would need another $200 every time someone wanted to use it . This way it will be accessible to more classes .
The cost to run it is a $ 27 Dye kit which lasts about 9 Pracs .

I made up some practice Dyes out of Bromothymol Blue powder and Methyl orange powder in a 20/80 glycerol /water mix . The kids can practice away with their 10uL micropipettes , befor we let them loose on the expensive Dyes :-o
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