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Lost cause??


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4 replies to this topic

#1 clarem

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Posted 20 January 2011 - 04:49 AM

So I did something really stupid and left some parts of a kit at room temperature, when they should have been stored at -20C (I know I'm an eejit :unsure: ). The kit arrived in a box with ice packs, and parts of it were left in the box under the ice packs for about 2 weeks.
The kit in question is the Biosciences RT2 Profiler PCR Array System, and the components which were left at room temperature include an RT cocktail (which includes an RT enzyme for conversion of RNA into cDNA), and a qPCR master mix (which includes SYBR Green).

This all happened over the summer and now the time has come where I need to use the kit. I am also quite limited with the amount of time I have left to do experiments.
So I am just wondering what peoples opinions are on the viability of this kit? Any chance it would work despite my stupid mistake?
Would it be a complete waste of time trying out the kit to see if it works anyway? Or since I'm quite time limited, should I just order a new kit straight away and forget about wasting time testing the old one(money is the issue here, it's quite an expensive kit).

I'd really appreciate any thoughts anyone has on this :)

#2 UBClabbie

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Posted 20 January 2011 - 12:35 PM

I would just order another kit. Enzymes don't do very well when left out at room temperature, especially for that extended period of time.
Used that kit once, and if i recall correctly u do not need to use their reverse transcription kit to use the array. Call and check because you can purchase RT kits for cheaper elsewhere ie. www.abmgood.com

definately order new mastermix

#3 clarem

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Posted 21 January 2011 - 04:09 AM

I would just order another kit. Enzymes don't do very well when left out at room temperature, especially for that extended period of time.
Used that kit once, and if i recall correctly u do not need to use their reverse transcription kit to use the array. Call and check because you can purchase RT kits for cheaper elsewhere ie. www.abmgood.com

definately order new mastermix



Thanks for your reply. I have just gone ahead and ordered a fresh kit, it probably is for the best.
Eek I definitely won't be making that mistake again :rolleyes:

#4 UBClabbie

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Posted 21 January 2011 - 09:49 AM

ya :( hope it works out for you.
even mastermix can go bad when stored at -20C for too long... some last 1 month, some only a month....
if you aren't going to use all your mastermix at once, aliquot it out and re-freeze. freeze thawing once should be ok, but don't do it more than that or you will comprimise the enzyme

#5 Trof

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Posted 31 January 2011 - 07:55 AM

I once forgot purified plasmids in the corner of the workbench and found them month later. They were to be send overseas dryied on a Whatman paper and they got there just fine. I guess that buffered circular DNA could survive anthing.

Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.

I never trust anything that can't be doubted.

'Normal' is a dryer setting. - Elizabeth Moon





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