New England Biolabs, E5530S, NEBNext® Single Cell Lysis Module

Related Categories RNA Library Prep for Illumina,, Next Generation Sequencing Library Preparation Specification Materials Required but not Supplied Nuclease-free Water DNA LoBind Tubes (Eppendorf® #022431021) Thermocycler Microcentrifuge FAQ Q: What cell types does the NEBNext Single Cell Lysis Module (NEB #E5530) support? A: The module is optimized for use with a variety of eukaryotic cell types. However, the cell lysis is not compatible with fixed cells or cells with cell walls (e.g., plant and fungal cells). Q: Can I resuspend the cells in buffers/medium other than PBS before cell collection? A: We strongly recommend that cells be resuspended in PBS buffer before cell collection. If PBS alone is not a viable option, cells can be resuspended in PBS supplemented with BSA. Up to 0.5 ng of BSA can be included in the cDNA Synthesis and Amplification reaction. If cells are resuspended in cell culture medium, for example, DMEM with 10% FBS, the carry-over volume during cell collection should be minimal (10-15 nl). For sorted cells do not use more than 5 µl carryover volume. Please see this FAQ. Q: Do the cells have to be sorted in to a specific reagent? A: We recommend that the cells be sorted into 1X Cell Lysis Buffer as instructed in the manual. Q: Can frozen cells be used? A: Yes. The cells can be sorted into 1X Cell Lysis Buffer, flash frozen and stored at -80°C. If, instead, the cells are collected in PBS or water before freezing, make the 1X Cell Lysis Buffer according to Section 1.2 (Cell Collection and Lysis) of the manuals supplied with the NEBNext Single Cell/Low Input RNA Library Prep Kit (NEB #E6420) or the NEBNext Single Cell/Low Input cDNA Synthesis & Amplification Module (NEB #E6421), taking account of the volume the cells are stored in.