Hola a Todos!!
We are working with Truseq Kit. with an input of 100 ng.
The samples are cellular culture supernatant and have amplified with primers Random. The quantification is done by Qubit.
The samples were pooled together in 4-5 samples with equimolar amounts and RSB elute (Tru Seq Illumina Kit) and fragment into Bioruptor and qc in this way we do it Bionalizador.
We have continued with the procedures as it indicates the Guide (end repair, adenilation, ligation and enrichment (PCR) with steps with Beads purification kit)
We qc selection to the size where we see our data, but finally when the PCR and purification is performed, bionalizador and took him to the library is lost.
You know if anyone has had this similar problem or that might be happening, and we are following the guidance indicating and maintaining our cold chain at all stages.
I wait your answer
We are working with Truseq Kit. with an input of 100 ng.
The samples are cellular culture supernatant and have amplified with primers Random. The quantification is done by Qubit.
The samples were pooled together in 4-5 samples with equimolar amounts and RSB elute (Tru Seq Illumina Kit) and fragment into Bioruptor and qc in this way we do it Bionalizador.
We have continued with the procedures as it indicates the Guide (end repair, adenilation, ligation and enrichment (PCR) with steps with Beads purification kit)
We qc selection to the size where we see our data, but finally when the PCR and purification is performed, bionalizador and took him to the library is lost.
You know if anyone has had this similar problem or that might be happening, and we are following the guidance indicating and maintaining our cold chain at all stages.
I wait your answer