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  • 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?

  • #2
    We had no problems when re-freezing HiSeq reagents once.

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    • #3
      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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      • #4
        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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        • #5
          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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          • #6
            @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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            • #7
              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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              • #8
                I suppose it will work with nextseq cartridges as well? I accidentally thawed the wrong one and refroze it. Now I would like to use it hoping it will work. Anyone has tried before?

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                • #9
                  @thiNGS, I accidentally thawed the wrong Nextseq 500 reagent cartridges and refrozen it. I would like to know about your result. Is it worked fine?

                  Thank you
                  Saravanan

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                  • #10
                    Hi! Yes it did work

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                    • #11
                      Thank you so much for your reply.

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