Cell Line Protocols
These protocols include directions for preparing medium and maintaining the cell lines, along with a few protocols for experimental procedures using cell lines. We expect to add additional protocols as we find time to do so. In addition, users who are not experienced with Drosophila cell culture may find it useful to read on of these reviews.
The following protocols are available:
- General Procedures for Maintenance of Drosophila Cell Lines - general overview of methods for growing the lines
- Maintenance of the ovarian line fGS/OSS - specific instructions for this line.
- Tissue Culture Medium - brief survey of the media that we commonly use
- Additions to Tissue Culture Medium - information about fetal calf serum, insulin, fly extract, and selective agents
- Thawing Cells - What to do with the frozen ampoule that we have shipped to you.
- Freezing Cells - How to prepare a stable frozen stock from a cell line.
- Immunostaining of Drosophila cells - Kris Kluegâs protocol (old but effective)
- Gelutol - Preparation of gelutol mounting medium for use in the immunostaining protocol.
- Creating new embryonic cell lines - Detailed description of the procedure used in the Miyake laboratory for generating embryonic cell lines in the 1980s. This protocol was written with the help of Ryu Ueda and Kumiko Ui-Tei, who made many of the cell lines in our collection when they were post-docs in the Miyake laboratory.
- Sources of Cell Culture Supplies - Where the DGRC buys chemicals and plasticware for cell culture.
- Medium for OSS Cell - Protocol for making medium for the ovarian lines OSS and fGS/OSS.
- RNA extraction from cells: modENCODE protocol #1 - Protocol used for preparation of RNA samples by modENCODE. This procedure, which does not preserve small RNAs (e.g. miRNAs), was used for most modENCODE transcriptome analyses.
- RNA extraction from cells: modENCODE protocol #2 - Protocol used for preparation of RNA samples by modENCODE when preservation of small RNAs (e.g. miRNAs) was required.
- Transferring cells using trypsinization - Trypsinization procedure that we recommend for a few strongly surface-adherent cell lines. Most of our cell lines can be transferred easily without trypsin treatment.
- Thawing, Culturing, and Cryopreserving Drosophila Cell Lines - Protocols for handling Drosophila cell lines. A video tutorial is also available.