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First ever project suggestions? - Starting genetic lab for hobby purposes.


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#16 pito

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Posted 28 June 2015 - 09:11 AM

a -22°C freezer is ok.

It will be ok for shorter term storage of competent cells, but I guess you are not going to store huge amounts of them for a long time.

 

Forget the dry ice, thats too expensive for long term storage, you could use it for a short term, but -22°C will do as well.


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#17 Dr. N00b

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Posted 28 June 2015 - 09:18 AM

a -22°C freezer is ok.

It will be ok for shorter term storage of competent cells, but I guess you are not going to store huge amounts of them for a long time.

 

Forget the dry ice, thats too expensive for long term storage, you could use it for a short term, but -22°C will do as well.

Phew! That really saved my day. However... please define "short term"? Is that like... 2 weeks, 2 months, 6 months? what? :)


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#18 pito

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Posted 28 June 2015 - 10:19 AM

hard to tell..

 

Most people keep competent cells in -80°C freezers, however some keep them in -22°C, but for how long? I do not know.

In most labs they just put them in the -80°C...

 

THe general idea or rule is to store them in -80°C freezers (or lower) and most people do not wonder about "what if". However: I know some keep them in -20°C and it worked.

Maybe the efficiency will be lower, but for your this is better than nothing at all because a -80°C seems to be out of the question and working with dry ice is also not really a possibility for long term storage.

 

Perhaps this can be a first experiment for you: making cells competent, transform them with a plasmid and store the rest of the cells in the -22°C and redo the experiment every week to see how good it works...

You can even try out different ways to make cells competent! This is even something that can be published! A lot of fuzz is still going around on competent cells and how to prepare them!


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#19 Dr. N00b

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Posted 28 June 2015 - 11:02 AM

Perhaps this can be a first experiment for you: making cells competent, transform them with a plasmid and store the rest of the cells in the -22°C and redo the experiment every week to see how good it works...

You can even try out different ways to make cells competent! This is even something that can be published! A lot of fuzz is still going around on competent cells and how to prepare them!

 

Great idea! laugh.png I will totally do that...


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#20 pito

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Posted 28 June 2015 - 11:20 AM

http://schneider.nci.../TIBS/feb96.txt

you can start with this link, to find out what method you would prefer. I also attached two papers mentiond in that link. So you can , for example, pick one of those methods to prepare your competent cells.

 

It could be a nice experiment to perhaps try both methods (make cells competent with both methods), then transform with a plasmid and check for the efficiency.

And store competent the cells in -22°C  and redo the transformation every week and check how the efficiency is influenced by the storage time...

 

This is just a simple project (also pretty cheap) to start with.

 

Perhaps you can do it with more than 1 type of cells: eg. E. coli top10, E.coli dh5alpha, and...

 

 

it could already be a nice experiment and its pretty easy and cheap to try/do.

 

All you need are the cells and the plasmid!

(and the reagents of course)

Attached Files


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#21 Dr. N00b

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Posted 28 June 2015 - 01:15 PM

http://schneider.ncifcrf.gov/methods/TIBS/feb96.txt

you can start with this link, to find out what method you would prefer. I also attached two papers mentiond in that link. So you can , for example, pick one of those methods to prepare your competent cells.

 

It could be a nice experiment to perhaps try both methods (make cells competent with both methods), then transform with a plasmid and check for the efficiency.

And store competent the cells in -22°C  and redo the transformation every week and check how the efficiency is influenced by the storage time...

 

This is just a simple project (also pretty cheap) to start with.

 

Perhaps you can do it with more than 1 type of cells: eg. E. coli top10, E.coli dh5alpha, and...

 

 

it could already be a nice experiment and its pretty easy and cheap to try/do.

 

All you need are the cells and the plasmid!

(and the reagents of course)

 

Thank you.

 

However, both these protocols require a refrigerated microcentrifuge. So I guess I need to get one of those as well then?


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#22 pito

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Posted 28 June 2015 - 11:11 PM

eum yes, I was assuming you had one.

In many protocols you do need this....

 

well, I guess it is something you really need if you want to do some microbiology. On the other hand: try it with a regular one and see what happens? I actually wonder what would happen. I wonder if it would still work at lower efficiency or not work at all!

 

Anyway: yeah, a cooled centrifuge is something you do need, but its expensive! (you are not really picking a cheap hobby lol)


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#23 bob1

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Posted 29 June 2015 - 12:15 AM

If you can get hold of a regular centrifuge, and have a bit of know-how you can put it inside a fridge and keep it cold that way. Just don't go putting it in and taking it out, as it will get condensation on the circuitry, which will fry the electronics when you go to use it.



#24 pito

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Posted 29 June 2015 - 01:07 AM

He is from Norway, maybe he can just put the centrifuge outside... haha

 

I dunno whether putting it inside a fridge would be a problem or not. Maybe not if you keep it permanently in it? I am assuming the centrifuges are build to cope with this?


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#25 Dr. N00b

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Posted 29 June 2015 - 02:41 AM

Damn... When I bought my current microcentrifuge, I kind of chose a non refridgerated one over a refridgerated one because of the price. And now it seems I need to buy one anyways, which will basically render my current one useless. :(


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#26 pito

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Posted 29 June 2015 - 03:27 AM

yes: its indeed better to buy one that can be cooled rather than 2.

 

Well, something you learned again!

 

I do not know, perhaps putting it in your refrigerator might work as well. I never tried it, but who knows?


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#27 Dr. N00b

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Posted 29 June 2015 - 03:44 AM

My rotor can keep 0.5ml, 1.5ml and 2ml tubes. It has two rows, one 0.5ml and one 2ml. I wonder.. What if I put the whole damn rotor in the freezer... And also freeze up a whole bunch of 2ml tubes with water. If taking the rotor out and placing in my centrifuge and use it right away, it should be able to hold a low temperature I think. Not exactly 4C, but maybe in the range 0 - whatever.

 

Does the temperature need to be 4 C exactly, or? ...


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#28 pito

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Posted 29 June 2015 - 03:51 AM

It gets hot pretty fast when centrifuging... Keep in mind: you often have to centrifuge for 10 minutes (or longer) so working with something you cooled in the freezer and then use at room temperatures will pretty much mean you end up at room temperatures very fast!

You are centrifuging at high speeds, generating lots of energy.


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#29 Dr. N00b

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Posted 29 June 2015 - 05:56 AM

Damn... I paid like 600 USD for my non refrigerated one. I see a couple of refrigerated units on e-bay for 800 USD a piece. Then there's shipping, and 25% VAT on top of that.

 

I need to sell my current one before I can make a purchase then. Unless I can use a cooling element from a mini refrigerator or whatnot and hack it onto the centrifuge. Maybe go in through the lid on the top, having it blow cool air through the bastard...

 

Peltier cooling element for CPU fans comes to mind...


Edited by Dr. N00b, 29 June 2015 - 05:57 AM.

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#30 pito

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Posted 29 June 2015 - 06:07 AM

well this is more a question for people in the technical field haha.

Perhaps you can just run it in a refrigerator? I guess you can brows the net a bit and find a forum where you could discuss this.


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