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Old 05-03-2013, 10:53 AM   #1
microgirl123
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Default Expired MiSeq reagent kit

Has anyone tried to run an expired MiSeq reagent kit? I've been working on developing an amplicon sequencing protocol, and the libraries look good through the KAPA quantification. Now I want to test them on the MiSeq without spending $1000 on a kit. We have an expired kit that we can't use for an actual, paying investigator so I wanted to at least try it out on my amplicons. Does anyone know if there's an expiration date built into the RFID tag? Our FAS didn't know.
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Old 05-03-2013, 11:05 AM   #2
kcchan
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I've tried using expired kits before and the instrument does let you run them, but the read quality does seem to be a bit worse than a new kit. If you have nothing to lose other than a few hours of your time, I would just try it and see how things go.
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Old 05-03-2013, 11:07 AM   #3
microgirl123
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Thanks! It's super expired (back in December), but I don't have anything to lose but time and if this method works, it should be a much cheaper way to sequence amplicons than the Caporaso, multiple-primer approach.
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Old 05-03-2013, 01:40 PM   #4
Harlon
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We've run expired kits before (always on non-critical samples), and they have all worked for us. I can't speak to the read quality - never bothered to check.

h
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Old 05-07-2013, 07:06 AM   #5
K.Hopkins
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We have run a couple of kits roughly 6 months out of date. The cluster density was a little low but the data quality was really good.

It would be good to know what concentration you load and what cluster density you get as we have another expired kit waiting for trial samples to be run on it.
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Old 05-08-2013, 03:54 AM   #6
matth431
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Our kits ran out in August 2012 and are still working fine, just click "Proceed anyway" when challenged on the RFID.
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Old 05-08-2013, 06:35 AM   #7
ZAAB
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Default Agreed

We've run quite a few 'expired' kits...

Never had a problem at all...Data Look great.

We cut anything with a Qscore <30, though...but that yield is still ~85% of total reads.

SInce the kits are shipped on dry ice, we store our kits at -80 for bulk storage, and I bring about 4 at a time into a -20...easier to access, and a bit faster to thaw out. I figure that the reagents are fine at -80 (in reality, -70).
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Old 05-13-2013, 05:45 AM   #8
microgirl123
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So I ran the expired kit (about 5 months out of date). The data looks great. The cluster density was a little low (I was aiming for 850K and got 630K), but that may have been because the titer of the phiX that I have is low and I used a 50% spike.
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Old 03-10-2016, 08:09 PM   #9
sdbrown
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sorry to dredge up an old thread but i wanted to tack on another data point.

this week i ran a 1-2 month expired (chip was 1 month, reagents were 2 months somehow) MiSeq v3 150-cycle kit and it worked beautifully despite the dire warning screen i had to click through.
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Old 03-22-2016, 11:18 AM   #10
mgangel
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Default And another data point

We routinely use OLD V3 kits. The reagent cartridge is stored @ -80C and the flow cell @ 4C. The oldest I've used is about 8 months expired without issue or significant degradation.
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Old 03-31-2016, 08:00 AM   #11
Ancestro
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They still can be used, even for months. If a year since expiration has passed, just reduce the cycle numbers to the 2/3 of the cap e.g. 600cycles to 400 for run.
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Old 04-01-2016, 05:43 AM   #12
dannyhi321
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Quote:
Originally Posted by Ancestro View Post
They still can be used, even for months. If a year since expiration has passed, just reduce the cycle numbers to the 2/3 of the cap e.g. 600cycles to 400 for run.
As long as they are not LESS than a year of course (V3 600)!
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Old 11-21-2016, 01:23 PM   #13
pedrolcalves
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Quick question,

For 300 cycles, how many different sequencing reactions would the Miseq Reagent Kit v2 be able to do?
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Old 11-21-2016, 04:45 PM   #14
nucacidhunter
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I hope this is what you are asking. Following is MiSeq v2 kit size and max number of cycles:
50 kit: 75 cycles
300 kit: 325 cycles
500 kit: 525

Note: if doing dual index sequencing, 7 dark cycles need to be accounted as well.
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Old 11-23-2016, 02:56 PM   #15
pedrolcalves
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Thank you for your reply. My main concern is whether the reagent kit serves only a single run or else, how much sequencing runs (different/independent) will I be able to do with the purchase of a Miseq reagent kit v2. Does the reagent cartridge serve a single sequencing run only? I'm sorry if I sound a little detached from reality. It's just that I'm learning about Illumina solutions as of now (for a theoretical project) and I lack lab experience regarding NGS.
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Old 11-23-2016, 03:09 PM   #16
nucacidhunter
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Each MiSeq sequencing kit is for one run only and you can multiplex several libraries in one run depending on number of required reads. Multiplexing can be tailored to obtain equal or unequal number of reads for each library.
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Old 12-07-2016, 07:08 PM   #17
GA-J
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Default NextSeq kit close to expire date

Does anybody know the different between NextSeq High output 300cycles and 150cycles kit? Only the reagent volume different? May I combine 2 of 150cycles kit for a 300 cycles run?
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Old 12-08-2016, 04:22 AM   #18
kmcarr
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Quote:
Originally Posted by GA-J View Post
Does anybody know the different between NextSeq High output 300cycles and 150cycles kit? Only the reagent volume different? May I combine 2 of 150cycles kit for a 300 cycles run?
No, you couldn't. The NextSeq, like the MiSeq, has all reagents provided in a single slide-in cartridge. The instrument will read the barcode on the reagent cartridge and determine from that the number of cycles which that cartridge can be used for. There is no way to override this.

[Disclaimer: My answer is not based on first hand knowledge but on assuming that the NextSeq operates like the MiSeq in this respect.]
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