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Old 04-23-2015, 07:45 AM   #1
jov14
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Default refreeze thawed reagent cartridge for Miseq (v3)

Hi everyone,
We accidentally thawed the wrong type of reagent cartrige for loading samples onto our Illumina MiSeq machine. We noticed only after the cartrige was completely thawed, that it contained reagents for only 152 cycles, whereas we needed 600 cycles.
Of course we do not want to throw the reagent cartridge away, so we opted to refreeze it for now.
I know that Illumina does not recommend using refrozen reagent cartridges, but I do not see why it should not work...

Does anybody have any experience with MiSeq sequencing runs using a reagent cartridge that was already thawed once and then refrozen?
Or does anybody know specific reasons why one should not try to use refrozen reagent cartridges?
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Old 04-23-2015, 11:29 AM   #2
luc
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We had no problems when re-freezing HiSeq reagents once.
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Old 04-24-2015, 12:51 AM   #3
dfhdfh
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Quote:
Originally Posted by jov14 View Post
Or does anybody know specific reasons why one should not try to use refrozen reagent cartridges?
It's probably for the same reason the food industry says you shouldn't refreeze thawed frozen food: When you throw it away and buy new food, they'll get more money.

In molecular biology that comparison might of course be a bit lackluster, however, we didn't have any trouble using a refrozen v2 50 cycle kit.
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Old 04-24-2015, 02:12 AM   #4
jov14
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Ok, thanks. It's a great relief to know that this cartridge is not clearly lost. It'll be a while till we may use this specific (150 cycle) cartridge, but when we do, I'll try to remember to post here how it went for us.
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Old 01-05-2016, 06:14 PM   #5
carladosanjos
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Default Results?

jov14, please let me know if your run has worked. Have you got an acceptable cluster density and quality?

I've thawed a 600 cycle v3 kit yesterday and I will need to postpone the sequencing due to a power outage. I am considering to refreeze the kit, but first I want to hear a bit about your results.

Cheers
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Old 01-07-2016, 12:21 AM   #6
jov14
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@carladosanjos: It worked fine. The quality was not worse than for any normal run. It's definitevely worth giving it a shot instead of throwing away the cartridge.
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Old 01-25-2016, 09:12 AM   #7
gwilkie
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Illumina state you can keep a thawed MiSeq eagent cartridge in the fridge for up to 7 days before using it. We have done this previously with no problems. Might be better than freeze/thaw if you are planning to use it within a week.
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