Thermo Scientific NGS library preparation, cleanup and size-selection products combine all necessary NGS library construction steps into one streamlined user-friendly workflow. Optimized for maximal performance and flexibility Thermo Scientific reagents allow for fast, effortless and reliable DNA library construction from minimal DNA sample input.
The Thermo Scientific product portfolio provides two distinct NGS library construction technologies – ClaSeek and MuSeek – each created to meet researcher's most demanding needs. The unmatched versatility of ClaSeek kits and incredible ease of use of MuSeek kits enables scientists to achieve excellent results with less effort and more time saved.
| ClaSeek | MuSeek | |
|---|---|---|
| Sample input | 5 ng-1 µg physically sheared DNA | 100 ng of intact genomic DNA, cDNA |
| Method | Ligation-based | Transposon-based |
| PCR-free protocol available | Yes | No |
| Physical sample fragmentation required | Yes | No (transposon-mediated) |
| Protocol | <60 min PCR-free protocol ~120 min amplification protocol | <80 min total time |
| Barcoding compatibility | Yes | Yes |
| NGS platform compatibility | Illumina MiSeq©, HiSeq, Ion Torrent PGM©, Ion Proton© | Ion Torrent PGM©, Ion Proton©, Illumina MiSeq™ Illumina HiSeq™ |
Unmatched Flexibility
Thermo Scientific ClaSeek Library Preparation Kits are designed for fast, convenient and accurate DNA library construction from DNA sample input as low as 5 ng. Available amplification-free protocols eliminate any possible PCR-related bias, resulting in uniform data coverage and reliable sequencing results.
Features:
Speed and convenience
Thermo Scientific MuSeek library preparation kits utilize an innovative transposon-based technology that allows enzymatic fragmentation and simultaneous tagging of the target DNA, eliminating the need for physical shearing, DNA end-repair, and adapter ligation steps. This enables fast library construction with minimal hands-on time and less effort.
Features:
MuSeek Library Preparation Kits utilize an innovative transposon-based technology that allows enzymatic fragmentation and simultaneous tagging of the target DNA, eliminating the need for physical sample shearing, DNA end-repair, and adaptor ligation steps.
The fragmentation and tagging reaction is catalyzed by a homotetrameric MuA transposase complex (75 kDa protein originating from bacteriophage Mu) containing two 50 bp double-stranded transposon DNA ends with a specific MuA binding sequence. Initially, the genomic DNA sample is simultaneously fragmented and tagged with transposon sequences at each end of resulting DNA fragments (1). In the next step the 3'-ends of the fragmented DNA are elongated over the 5 nt gaps generated during the transposition reaction and further extended into transposon sequences (2) . Sequencing platform-specific adaptors are added to the fragments in a subsequent PCR-based adaptor-addition reaction. In the initial cycles of PCR the first 16 nt of the adaptor 3'-ends hybridize to transposon sequences in tagged DNA fragments (2). In later PCR cycles the fragments are amplified using a pair of external primers.
Reliable NGS Library Cleanup and Size Selection
Thermo Scientific MagJET and GeneJET NGS Cleanup and Size Selection kits are optimized for robust and efficient DNA fragment library purification and size selection from a large variety of enzymatic reaction mixtures used in NGS workflows.
Features:
Fragmented E. coli genomic DNA (1 µg) samples, each spiked in different reaction mixtures and buffers (a-h) typical for next-generation sequencing workflow, were size selected using MagJET and other supplier protocols. Isolated fragment libraries were analyzed on an agarose gel. Expected size selection peak areas are marked with a dotted line.
a – End Conversion Reaction mixture, IT;
b – PCR Reaction mixture (1);
c – PCR Reaction mixture (2);
d – Ligation Mixture;
e – End Conversion Reaction mixture (IL);
f – Fragmentation reaction buffer;
g – Elution buffer;
h – Nebulization buffer;
C – Control (input DNA)
