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Author: Subject: DIY Centrifuge?
mayko
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[*] posted on 5-7-2020 at 15:36


Another Prakash Lab invention: the HandyFuge, based on the principle of a squeeze-charge flashlight

Handyfuge-LAMP: low-cost and electricity-free centrifugation for isothermal SARS-CoV-2 detection in saliva.
Ethan Li, Adam Larson, Anestha Kothari, Manu Prakash
medRxiv 2020.06.30.20143255; doi: https://doi.org/10.1101/2020.06.30.20143255

Quote:

Point of care diagnostics for COVID-19 detection are vital to assess infection quickly and at the source so appropriate measures can be taken. The loop-mediated isothermal amplification (LAMP) assay has proven to be a reliable and simple protocol that can detect small amounts of viral RNA in patient samples (<10 genomes per μL) (Nagamine, Hase, and Notomi 2002) Recently, Rabe and Cepko at Harvard published a sensitive and simple protocol for COVID-19 RNA detection in saliva using an optimized LAMP assay (Rabe and Cepko, 2020). This LAMP protocol has the benefits of being simple, requiring no specialized equipment; rapid, requiring less than an hour from sample collection to readout; and cheap, costing around $1 per reaction using commercial reagents. The pH based colorimetric readout also leaves little ambiguity and is intuitive. However, a shortfall in many nucleic acid-based methods for detection in saliva samples has been the variability in output due to the presence of inhibitory substances in saliva. Centrifugation to separate the reaction inhibitors from inactivated sample was shown to be an effective way to ensure reliable LAMP amplification. However, a centrifuge capable of safely achieving the necessary speeds of 2000 RPM for several minutes often costs hundreds of dollars and requires a power supply. We present here an open hardware solution- Handyfuge - that can be assembled with readily available components for the cost of <5 dollars a unit and could be used together with the LAMP assay for point of care detection of COVID-19 RNA from saliva. The device is then validated using the LAMP protocol from Rabe and Cepko. With the use of insulated coolers for reagent supply chain and delivery, the assay presented can be completed without the need for electricity or any laboratory scale infrastructure.




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Ubya
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[*] posted on 5-7-2020 at 17:57


Quote: Originally posted by mayko  
Another Prakash Lab invention: the HandyFuge, based on the principle of a squeeze-charge flashlight

Handyfuge-LAMP: low-cost and electricity-free centrifugation for isothermal SARS-CoV-2 detection in saliva.
Ethan Li, Adam Larson, Anestha Kothari, Manu Prakash
medRxiv 2020.06.30.20143255; doi: https://doi.org/10.1101/2020.06.30.20143255

Quote:

Point of care diagnostics for COVID-19 detection are vital to assess infection quickly and at the source so appropriate measures can be taken. The loop-mediated isothermal amplification (LAMP) assay has proven to be a reliable and simple protocol that can detect small amounts of viral RNA in patient samples (<10 genomes per μL) (Nagamine, Hase, and Notomi 2002) Recently, Rabe and Cepko at Harvard published a sensitive and simple protocol for COVID-19 RNA detection in saliva using an optimized LAMP assay (Rabe and Cepko, 2020). This LAMP protocol has the benefits of being simple, requiring no specialized equipment; rapid, requiring less than an hour from sample collection to readout; and cheap, costing around $1 per reaction using commercial reagents. The pH based colorimetric readout also leaves little ambiguity and is intuitive. However, a shortfall in many nucleic acid-based methods for detection in saliva samples has been the variability in output due to the presence of inhibitory substances in saliva. Centrifugation to separate the reaction inhibitors from inactivated sample was shown to be an effective way to ensure reliable LAMP amplification. However, a centrifuge capable of safely achieving the necessary speeds of 2000 RPM for several minutes often costs hundreds of dollars and requires a power supply. We present here an open hardware solution- Handyfuge - that can be assembled with readily available components for the cost of <5 dollars a unit and could be used together with the LAMP assay for point of care detection of COVID-19 RNA from saliva. The device is then validated using the LAMP protocol from Rabe and Cepko. With the use of insulated coolers for reagent supply chain and delivery, the assay presented can be completed without the need for electricity or any laboratory scale infrastructure.


this is ok for 0.5-1.5ml tubes, i would not try this with my 15ml glass centrifuge tube





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Mateo_swe
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[*] posted on 7-7-2020 at 10:34


Hmm, i would like a centrifuge with 1000ml bottles.
I guess it could be done with some DIY experimentation.
I dont need any 1000 of rpms, i would guess bike wheel speed would be enough.
And at these speeds it would be easier to shield and a breakdown would only damage the centrifuge and samples.
It would be nice for hard to filtrate stuff.
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draculic acid69
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[*] posted on 9-7-2020 at 05:24


Quote: Originally posted by ScienceSquirrel  
Quote:

I've seen some designs on-line but none of them impressed me and many of them do not look safe. For example...
http://hackteria.org/wiki/index.php/DIY_handheld_centrifuge


That looks more like a weapon than a piece of scientific equipment. Imagine turning up at your local hospital with a piece of PVC tube embedded in your chest. :(


That thing looks dangerous.
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Tsjerk
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[*] posted on 9-7-2020 at 05:52


Quote: Originally posted by draculic acid69  
Quote: Originally posted by ScienceSquirrel  
Quote:

I've seen some designs on-line but none of them impressed me and many of them do not look safe. For example...
http://hackteria.org/wiki/index.php/DIY_handheld_centrifuge


That looks more like a weapon than a piece of scientific equipment. Imagine turning up at your local hospital with a piece of PVC tube embedded in your chest. :(


That thing looks dangerous.


I laughed at the PRECAUTIONS section :)
Quote:
....if possible use a protective shield, ex: a riding helmet......
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draculic acid69
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[*] posted on 9-7-2020 at 17:26




I laughed at the PRECAUTIONS section :)
Quote:
....if possible use a protective shield, ex: a riding helmet......
[/rquote]


Yeah that'll protect you when six weighted swinging pvc tubes spinning at a few thousand Rpm's smack into your arm,body,neck,head & face

[Edited on 10-7-2020 by draculic acid69]
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[*] posted on 9-7-2020 at 19:23


Quote: Originally posted by Mateo_swe  
Hmm, i would like a centrifuge with 1000ml bottles.
I guess it could be done with some DIY experimentation.
I dont need any 1000 of rpms, i would guess bike wheel speed would be enough.
And at these speeds it would be easier to shield and a breakdown would only damage the centrifuge and samples.
It would be nice for hard to filtrate stuff.


How close do you live to a playground?



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SWIM
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[*] posted on 9-7-2020 at 20:02


Cream separators, anybody?

A nice hand -cranked DeLaval #18 maybe?




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draculic acid69
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[*] posted on 9-7-2020 at 21:56


In Australia we used to have hills hoist.it has a hand crank halfway up the middle pole.
One could attach many coke bottles or a few 20litre drums with some reinforcement if desperate/stupid enough to think it's a good idea

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Mateo_swe
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[*] posted on 10-7-2020 at 11:36


Something like this looks interesting.
It would need a sturdy shield.

And the playarea for children where i live dont have a merry-go-round (thats what they are called, right?)


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SWIM
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[*] posted on 10-7-2020 at 12:21


Quote: Originally posted by draculic acid69  
In Australia we used to have hills hoist.it has a hand crank halfway up the middle pole.
One could attach many coke bottles or a few 20litre drums with some reinforcement if desperate/stupid enough to think it's a good idea


Don't sell the hills hoist short.
They were all over US suburbs in the 60s and 70s too.

Some housing developments mandated their use because they didn't want clotheslines hanging all over the place.
They probably sold hundreds of thousands here. Maybe a million.

Gee, I wonder if spinning it like a centrifuge would save much time on drying your clothes?
I suppose your socks might get a little longer with time though.




Ebay says they need to get their hands on my bank account if I want to keep selling there.
This sounds like the best idea since putting ortho tricresyl phosphate in Ginger Jake.
I'm walking while I can still walk straight.




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draculic acid69
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[*] posted on 10-7-2020 at 17:36



Don't sell the hills hoist short.
They were all over US suburbs in the 60s and 70s too.

Some housing developments mandated their use because they didn't want clotheslines hanging all over the place.
They probably sold hundreds of thousands here. Maybe a million.

Gee, I wonder if spinning it like a centrifuge would save much time on drying your clothes?
I suppose your socks might get a little longer with time though.[/rquote]

I'm wondering how much weight you can hang off one before it goes bad?
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