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Tebubio's blog - Acting and reacting in life sciences and biotechnologies
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Gene Expression - Molecular Biology

Prevent adapter dimer formation during NGS library prep

10/11/2014 by Mark Livingstone 6 Comments

from: Shore et al. Improved Small RNA Library Preparation Workflows for Next Generation Sequencing [Poster]

Sources: Shore et al. “Improved Small RNA Library Preparation Workflows for Next Generation Sequencing” [Poster]

It is now quite easy to prevent adapter dimer formation during NGS library preps by using chemically-modified adapters.

In a poster presented at the recent ASHG 2014 meeting in San Diego, researchers from TriLink Biotechnologies described their innovative technology solving the adapter dimer problem in Next Generation Sequencing library prep.

Similar to the primer dimer problem observed when performing PCR (i.e. low molecular weight PCR by-products of primer-primer hybridization), adapter dimerization generates by products of NGS library prep that will greatly decrease the number of informative sequencing reads. This is particularly a problem for low-input studies and when working with small RNAs (e.g. miRNA).

TriLink’s expertise in chemistry-based solutions for biological problems has previously led to the creation of the CleanAmp™ product line. CleanAmp™ Primers, CleanAmp™ dNTPs and 2X PCR Master Mix employ heat labile modifications, similar to those on hot-start polymerases, to prevent primer dimerization or premature polymerase activity prior to a high-temperature activation step.

CleanAmp™ 7-deaza-dGTP is a chemistry-based solution allowing for the amplification of difficult GC-rich PCR products.

Interested in learning more about TriLink’s products and services (including GMP production of modified oligonucleotides for Phase I human trials)? Contact tebu-bio experts who are representing TriLink in Europe.

Gene Expression - Molecular Biology

The empirical approach to rRNA depletion for NGS

15/10/2014 by Mark Livingstone No Comments

Ribosomal RNA (rRNA) depletion is a must for Next Generation Sequencing (NGS) studies to reduce the number of irrelevant reads. While commercially-available kits such as the Ribo-zero or the BioMag® SelectaPure mRNA System are popular for rRNA depletion, there are some experimental situations in which another approach is required.

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