Focused Sequence Depth
The NEXTFLEX® Bisulfite-Seq Kit with reduced representation utilizes a restriction enzyme that leads to selective amplification of CpG regions, resulting in focused sequence depth. The NEXTFLEX® Msp1 Restriction Enzyme, which is available separately, has been optimized for reduced representation studies using the NEXTFLEX® Bisulfite-Seq Kit. The NEXTFLEX® Bisulfite-Seq Kit can also be used without the MSP 1 Restriction Enzyme for genome-wide methyl-seq analysis, including under-represented CpG regions.
KIT CONTENTS
- NEXTFLEX® Bisulfite-Seq End Repair Buffer Mix
- NEXTFLEX® Bisulfite-Seq End Repair Enzyme Mix
- NEXTFLEX® Bisulfite-Seq Adenylation Mix
- NEXTFLEX® Bisulfite-Seq Ligation Mix
- NEXTFLEX® Bisulfite-Seq DNA Adapter 24 (25 µM)
- NEXTFLEX® Adapter Dilution Buffer
- NEXTFLEX® Primer Mix (12.5 µM)
- NEXTFLEX® Bisulfite-Seq U+ PCR Master Mix
- Nuclease-free water
- Resuspension Buffer
REQUIRED MATERIALS NOT PROVIDED
- 10 ng – 1 µg of fragmented /or/ Msp1 digested genomic DNA in up to 40 µL nuclease-free water.
- NEXTFLEX® Bisulfite-Seq Barcodes – 6 / 12 / 24 (Cat # 511911, 511912, 511913 ) (Optional)
- NEXTFLEX® Msp1 – 8 reactions / 48 reactions (Cat # 511921, 511922)
- EZ DNA Methylation-Gold™ Kit (Zymo Research, Cat # D5005)
- Ethanol 100% (room temperature)
- Ethanol 80% (room temperature)
- 96 well PCR Plate Non-skirted (Phenix Research, Cat # MPS-499) / or / similar
- 96 well Library Storage and Pooling Plate (Fisher Scientific, Cat # AB-0765) / or / similar
- Adhesive PCR Plate Seal (BioRad, Cat # MSB1001)
- Agencourt AMPure XP 5 mL (Beckman Coulter Genomics, Cat # A63880)
- Magnetic Stand – 96 (Ambion, Cat # AM10027) / or / similar
- Heat block
- Thermocycler
- 2, 10, 20, 200 and 1000 µL pipettes / multichannel pipettes
- Nuclease-free barrier pipette tips
- Microcentrifuge
- 1.5 mL nuclease-free microcentrifuge tubes
- Vortex
