Rerun of cycle sequencing

mohd2b · 12-28-2015, 10:23 PM

Hi All

Recently I have run a cycle sequencing reaction, however the run was stopped due to a fatal error in the thermocycler. it stopped at cycle 8 of 25 cycles for approximately an hour till I realized the error message .then, I decided to rerun the cycle sequencing using different machine immediately.

from your experience , is the reaction is going to work if I sequence it. Should I stop and repeat the whole reaction again?

Thanks

MU Core · 12-29-2015, 07:25 AM

I assume you are using Big Dye v3 chemistry. I would expect the reaction to work as long as there were no other issues (i.e. primer is annealing, GC-rich template problems, etc.).

mohd2b · 12-29-2015, 08:54 PM

Hi MU Core

iam using BigDye v1.1 chemistry.

MU Core · 12-30-2015, 06:50 AM

Hello mohd2b,

There should be no problem completing the cycles and loading the reaction. We've had a similar issue with a 96 well plate and data looked fine. Good luck.

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12-28-2015, 10:23 PM	#1
mohd2b Junior Member Location: Oman Join Date: Nov 2014 Posts: 9	Rerun of cycle sequencing Hi All Recently I have run a cycle sequencing reaction, however the run was stopped due to a fatal error in the thermocycler. it stopped at cycle 8 of 25 cycles for approximately an hour till I realized the error message .then, I decided to rerun the cycle sequencing using different machine immediately. from your experience , is the reaction is going to work if I sequence it. Should I stop and repeat the whole reaction again? Thanks

12-29-2015, 07:25 AM	#2
MU Core Member Location: Columbia, Missouri Join Date: Apr 2008 Posts: 57	I assume you are using Big Dye v3 chemistry. I would expect the reaction to work as long as there were no other issues (i.e. primer is annealing, GC-rich template problems, etc.).

12-29-2015, 08:54 PM	#3
mohd2b Junior Member Location: Oman Join Date: Nov 2014 Posts: 9	Hi MU Core iam using BigDye v1.1 chemistry.

12-30-2015, 06:50 AM	#4
MU Core Member Location: Columbia, Missouri Join Date: Apr 2008 Posts: 57	Hello mohd2b, There should be no problem completing the cycles and loading the reaction. We've had a similar issue with a 96 well plate and data looked fine. Good luck.