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Thermo Scientific® MuSeek™ Library Preparation Kit for the Ion Torrent™ instrument - FAQs

查看更多产品信息 Thermo Scientific® MuSeek™ Library Preparation Kit for the Ion Torrent™ instrument - FAQs (4480829)

8 个常见问题解答

Is MuSeek suitable with ssDNA?

MuSeek is not suitable for ssDNA, as MuA transposase is not able to fragment ssDNA. To use MuSeek, ssDNA should first be converted into dsDNA.

Can I use MuSeek to construct NGS library from a PCR product?

Yes. However for construction of NGS library from PCR amplicons in fragmentation reaction, do not use PCR products shorter than 300 bp. Due to intrinsic features of the transposon technology, a approximately 50 bp drop off is expected in sequencing coverage from each distal end of the amplicon sequence. This can be averted by designing your amplicons to be approximately 100 bases larger than the desired sequencing insert.

Can I use lower than recommended sample amount for MuSeek fragmentation reaction?

No, the recommended sample DNA input should be followed strictly. The MuSeek fragmentation reaction is optimized to yield optimal fragment library lengths at recommended input.

Is MuSeek compatible with RNA library preparation workflow?

We do not supply specific MuSeek kits for NGS library preparation from RNA samples. However, MuSeek is compatible with double-stranded cDNA substrate.

What is the difference between the ClaSeek technology and MuSeek technology?

The ClaSeek technology uses a classical NGS library preparation approach where physically fragmented DNA is end-repaired and platform-specific adapters are added through ligation reaction. MuSeek transposase-based technology simultaneously fragments intact DNA and adds MuA transposon derived DNA sequence which is further used for PCR-based adapter addition step.

What is the sample input requirement for the MuSeek protocol?

MuSeek provides a protocol for NGS library construction from 50 ng or 100 ng sample DNA.

How does MuSeek DNA sample preparation technology work?

MuSeek technology utilizes the MuA transposase enzyme, which catalyzes simultaneous fragmentation of double-stranded target DNA and tagging the fragment ends with transposon DNA. In a subsequent PCR step, the platform-specific adaptors are added using Thermo Scientific Phusion High-Fidelity DNA polymerase.

What is the difference between EpiJET 5-hmC Analysis Kit (Cat. No. K1481) and EpiJET 5-hmC and 5-mC Analysis Kit (Cat. No. K1501)?

The new EpiJET 5-hmC and 5-mC Analysis Kit is more universal than the previous EpiJET 5-hmC Analysis Kit because it provides a total assessment of both cytosine modifications - 5-hydroxymethylcytosine (5-hmC) and methylcytosine (5-mC) - within specific DNA loci (for comparison - the current kit allows only 5-hmC analysis). Additionally, all necessary DNA controls do not originate from plasmids widely used in molecular biology laboratories are included into the new kit.