Description

Bisulfite Sequencing Library Prep Kit

 

The Bisulfite Sequencing Library Prep Kit (illumina platform) was developed for construction of NGS libraries using bisulfite treated DNA (50 ng – 500 ng) as input. DNA methylation is an epigenetic mechanism known to play a critical role in gene regulation and genomic imprinting by blocking transcription factor access to promoters and enhancers. Bisulfite sequencing is a popular technique in biomedical research based on C to T conversion under the treatment of sodium bisulfite.

Recently, NGS became a powerful tool to identify the DNA methylation status at the whole genome level with single-base resolution. However, it is well known that bisulfite treatment of the NGS libraries causes tremendous damage to the libraries.

In the case of the regular bisulfite seq library preparation (library prep before bisulfite conversion), the DNA shearing equipment and the expensive methylated adaptors are required. In addition, the subsequent bisulfite conversion causes tremendous DNA damage to the constructed libraries.

BioDynami has developed a unique library prep technology to solve the problems. The technology uses bisulfite treated DNA as input to avoid the significant library loss caused by bisulfite conversion. Furthermore, DNA shearing step and expensive methylated adaptors are not required with our kit. The DNA polymerase in the kit has high-fidelity amplification ability and uracil tolerance which is ideal for amplification of bisulfite sequencing libraries. The final library is strand specific.

 

Three index types are available for the kit:

Non-index (Cat.# 30091): Libraries do not have index.

Index (Cat.# 30092): Each primer contains a unique barcode sequence of 6 bases to identify the individual library. Library multiplexing capacity is up to 48 samples. Index information can be downloaded here.

Unique dual index (Cat.# 30093): The multiplexing of bisulfite sequencing library is up to 96 samples with unique dual indexes. We used a Four-Base Difference Index System to generate indexes that have at least 4 bases different from each other in the 8-base index. The index primers remove NGS errors including index cross-contamination, index hopping, reads mis-assignment etc. Index information can be downloaded here.

Kit advantages

• • • Easy and Quick protocol
      • Easy: Hands-on time only 10 minutes
      • Quick: Total protocol time around 1.5 hours
    • Simple workflow: Less steps
    • Directional library
    • Save magnetic beads more than 50%
    • Guaranteed high Bisulfite sequencing library conversion efficiency
    • Bisulfite treated DNA as input: From 50 ng to 500 ng

 

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