How much MiSeq sequencing reagent is being used per each cycle.

Mizraelson · 09-29-2017, 03:27 AM

Hi everyone. I have some thoughts to use leftovers after several MiSeq runs to perform one extra run (I do have an extra flowcell, but reagents are needed). So my question is how much reagent is used by MiSeq per each cycle? Does anyone now that or have an experience in using the rest of the reagent after a run?

GenoMax · 09-29-2017, 05:00 AM

I understand being frugal is a good thing but trying to re-use reagents that are several days old and stayed out at 6-8 degrees in the machine does not seem like a wise choice unless you want to do this purely as an experiment (you will need to pool reagents from multiple runs unless you have a large amount left over due to a short run with a long run kit).

Generally all kits have enough extra reagents to do 10-15 cycles (to account for index reads) as I recall.

Mizraelson · 09-29-2017, 06:17 AM

Sure, that's purely an experiment. Thats exactly what I want to do - to pool reagents from multiple runs and I do sometimes see there a lot left in the cartridge. So is there a way to more or less predict how much reagent (by volume) is needed for one cycle in order to extrapolate.

GenoMax · 09-29-2017, 07:15 AM

Someone with better experimental expertise than me will have to answer that.

kcchan · 09-29-2017, 12:34 PM

A lot of the leftover reagents is actually water from the template rinse and back flush. Probably not a good idea to re-use them.

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09-29-2017, 03:27 AM	#1
Mizraelson Junior Member Location: Moscow, Russia Join Date: Aug 2013 Posts: 3	How much MiSeq sequencing reagent is being used per each cycle. Hi everyone. I have some thoughts to use leftovers after several MiSeq runs to perform one extra run (I do have an extra flowcell, but reagents are needed). So my question is how much reagent is used by MiSeq per each cycle? Does anyone now that or have an experience in using the rest of the reagent after a run? Last edited by Mizraelson; 09-29-2017 at 03:39 AM.

09-29-2017, 05:00 AM	#2
GenoMax Senior Member Location: East Coast USA Join Date: Feb 2008 Posts: 7,080	I understand being frugal is a good thing but trying to re-use reagents that are several days old and stayed out at 6-8 degrees in the machine does not seem like a wise choice unless you want to do this purely as an experiment (you will need to pool reagents from multiple runs unless you have a large amount left over due to a short run with a long run kit). Generally all kits have enough extra reagents to do 10-15 cycles (to account for index reads) as I recall.

09-29-2017, 06:17 AM	#3
Mizraelson Junior Member Location: Moscow, Russia Join Date: Aug 2013 Posts: 3	Sure, that's purely an experiment. Thats exactly what I want to do - to pool reagents from multiple runs and I do sometimes see there a lot left in the cartridge. So is there a way to more or less predict how much reagent (by volume) is needed for one cycle in order to extrapolate.

09-29-2017, 07:15 AM	#4
GenoMax Senior Member Location: East Coast USA Join Date: Feb 2008 Posts: 7,080	Someone with better experimental expertise than me will have to answer that.

09-29-2017, 12:34 PM	#5
kcchan Senior Member Location: USA Join Date: Jul 2012 Posts: 184	A lot of the leftover reagents is actually water from the template rinse and back flush. Probably not a good idea to re-use them.