Header Leaderboard Ad

Collapse

NimbleGen SeqCap and empPCR lib L

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • NimbleGen SeqCap and empPCR lib L

    Dear all, I performed sequence capture of target exons using NimbleGen SeqCap EZ strategy. Now I have to proceed to emPCR (Lib L). Should I denature my for 95 C degrees for 2 minutes?

  • #2
    No you don't, after LM-PCR only 1 of the strands will be able to get PCRed on the bead.

    Comment


    • #3
      Ok, thank you Zaag. May I ask you how many copy per bead do you use in emPCR lib L? I would start trying 1 cpb since it is the first time that I perform this emPCR. Do you have any suggestion?

      Comment


      • #4
        After sequence capture I usually end up between 1 and 2 cpb; so if you measured your sample with a Qubit before calculating the nr of molecules my guess would be that 1cpb would be a safe number.

        Usually I do one SVE with 1 cpb and one with 2 cpb and after calculating enrichment I choose a cpb for my bulk emulsion.

        Comment


        • #5
          Can I ask you what is SVE?

          Comment


          • #6
            That would be a small volume emulsion. They come in three sizes, small medium and large.

            Comment


            • #7
              Ok.Thank you very much for your reply. I was wondering if you also perform SeqCap EZ Choice...I have a lot of question regarding this method but I don't think many people of this forum perform SeqCap and then run on a GS Junior platform

              Comment


              • #8
                Yeah we do that but on the 454-FLX+ so no junior here. I am not sure what exactly are the differences regarding emPCR and such, but if you have questions just mail me, or ask them here (mail is faster answer).

                Comment

                Latest Articles

                Collapse

                • seqadmin
                  A Brief Overview and Common Challenges in Single-cell Sequencing Analysis
                  by seqadmin


                  ​​​​​​The introduction of single-cell sequencing has advanced the ability to study cell-to-cell heterogeneity. Its use has improved our understanding of somatic mutations1, cell lineages2, cellular diversity and regulation3, and development in multicellular organisms4. Single-cell sequencing encompasses hundreds of techniques with different approaches to studying the genomes, transcriptomes, epigenomes, and other omics of individual cells. The analysis of single-cell sequencing data i...

                  01-24-2023, 01:19 PM
                • seqadmin
                  Introduction to Single-Cell Sequencing
                  by seqadmin
                  Single-cell sequencing is a technique used to investigate the genome, transcriptome, epigenome, and other omics of individual cells using high-throughput sequencing. This technology has provided many scientific breakthroughs and continues to be applied across many fields, including microbiology, oncology, immunology, neurobiology, precision medicine, and stem cell research.

                  The advancement of single-cell sequencing began in 2009 when Tang et al. investigated the single-cell transcriptomes
                  ...
                  01-09-2023, 03:10 PM

                ad_right_rmr

                Collapse
                Working...
                X