Hi all,
I am having problems with the NEBnext Ultra kit for library preparation. I´ve been doing same amplicon libraries for three years with success, but since October none of my libraries seems to hybridize with the adapters of the flow cell (I got 0 clusters). It happened already with 6 different runs (Miseq v3, 600 cycles) and 240 different library preps.
The bioanalyzer pattern was good (size OK, no adapters, shape OK), final pools were quantified by Qubit, Bionalyzer and qPCR (Qiaseq). None of the quantifications worked. PhiX was loaded at 10 and 15% equimolar to pooled libraries (12pM).
NaOH and Miseq are working because other researchers are using them with no problems.
Any idea of what can be the problem?. Neither Illumina, nor NEB, are helpful.
Thanks
I am having problems with the NEBnext Ultra kit for library preparation. I´ve been doing same amplicon libraries for three years with success, but since October none of my libraries seems to hybridize with the adapters of the flow cell (I got 0 clusters). It happened already with 6 different runs (Miseq v3, 600 cycles) and 240 different library preps.
The bioanalyzer pattern was good (size OK, no adapters, shape OK), final pools were quantified by Qubit, Bionalyzer and qPCR (Qiaseq). None of the quantifications worked. PhiX was loaded at 10 and 15% equimolar to pooled libraries (12pM).
NaOH and Miseq are working because other researchers are using them with no problems.
Any idea of what can be the problem?. Neither Illumina, nor NEB, are helpful.
Thanks
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