Diluted to 1X working solution (28 Wünsch units/vial)
RNase-AWAY
Sigma-Aldrich
83931
Biotinylated Rat anti-Mouse-CD8a antibody
Invitrogen
MCD0815
100 μg/ml stock concentration
BSA (Bovine Serum Albumin), Fraction V
GibcoBRL
11018-017
Prepare a 1% BSA solution in PBS
Dynabeads M-280 Streptavidin
Invitrogen
11205D
1 μl can bind 0.05-0.10 μg of biotinylated antibody
PicoPure RNA Isolation Kit
Molecular Devices
KIT0204
Follow manufacturer’s instructions
Equipment
(10) Neodymium block magnets (K & J Magnetics, Inc., Cat. #B444B), alternatively DynaMag™-2 (Invitrogen, Cat. #123-21D) may be used.
Kontes Glass Tissue Grinder, 2 ml working capacity, with large clearance pestle (Cat. #885300-0002)
Cell filters, e.g. MACS Pre-Separation Filter (Miltenyi Biotec, Cat. #130-041-407)
35 mm petri-dishes coated with Sylgard (Dow Corning Corporation, Cat. #3097358-1004)
Dissecting tools: two pairs of Dumont No. 5 forceps (Fine Science Tools, Cat. #11251-20)
Vannas spring micro-dissection scissors (Fine Science Tools, Cat. #15000-08)
Pipettes (P-1000, P-100, P-10) with disposable tips
Standard hemocytometer to count the cells and estimate purity
Pasteur pipettes with fire polished tips of standard diameter, narrowed to ~50% of standard diameter and narrowed to ~25% of standard diameter for trituration
Table top micro-centrifuge (1-16,000 (x) g)
Vortex
Fluorescent stereo-microscope (a Leica MZ16FA was used in this protocol)
Iyer, E. P. R., Iyer, S. C., Sulkowski, M. J., Cox, D. N. Isolation and Purification of Drosophila Peripheral Neurons by Magnetic Bead Sorting. J. Vis. Exp. (34), e1599, doi:10.3791/1599 (2009).