Pico Methyl-Seq Library Prep Kit
D5455 / D5456
- All-inclusive kit for bisulfite conversion followed by Whole Genome Bisulfite Sequencing (WGBS) library preparation.
- Accommodates ultra-low DNA input and compatible with FFPE samples.
- Simple, ligation- and gel-free workflow can be completed in a few hours.
The Pico Methyl-Seq Library Prep Kit provides a streamlined workflow for making WGBS libraries. Input DNA is randomly fragmented during the initial bisulfite treatment step followed by three rounds of amplification with uniquely designed primers. The procedure can accommodate as little as 10 pg input DNA (including that derived from FFPE samples), making it ideal for methylation analysis of precious, limited, and target-enriched samples.
|Input||10 pg - 100 ng|
|Sample Source||The protocol is designed for 10 pg – 100 ng genomic DNA input. DNA should be free of enzymatic inhibitors and can be suspended in water, TE, or a low salt buffer. DNA with low 260/280 or 260/230 ratios should be purified prior to processing using the Genomic DNA Clean & Concentrator (D4010).|
|Sequencing Compatibility||This kit produces libraries with Illumina TruSeq Adapters that can be sequenced on any of the Illumina sequencing platforms.|
Q1: Can cfDNA be used as an input?
Q2: Are there any rules for combining multiple index primers?
Q3: Are the index primers used in RRHP 5-hmC Library Prep kit (D5451) the same as in the Pico Methyl-Seq Library kit (D5455)?
Q4: How should sequenced Pico Mehtyl-Seq library be aligned?
Q5: Should single end or paired end read be performed? Which read length should be set?
Q6: What method of quantification is recommended for the input DNA? (PicoGreen, SYBR or maybe qPCR)?
Q7: What to do if the lid temperature of thermo-cycler does not go down to 25°C?
Q8: Why does the library banding pattern look like a DNA ladder?
Q9: I am working with only a few hundred cells. Can I use the EZ DNA Methylation – Direct kit for bisulfite conversion directly from cells?
Q10: What is the sequence of the PrepAmp Primer and the LibraryAmp Primers?
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