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Germ Cell Book
Protocols
EdU labeling and staining
Ballistic Transformation
Supplement
Dissection, staining, in situ
Gonad Dissections
Gonad Fixation
DAPI Staining
Antibody Staining of Dissected Gonads
Activated MPK-1 Staining (MAPK-YT, Diphospho-MPK-1)
Germline in situ hybridization
Worm Powder
RNAi stuff
RNAi plates with lactose (instead of IPTG)
Feeding RNAi
dsRNA (double-stranded RNA) for injection
Genomic stuff
Worm genomic DNA prep
Single worm PCR
Mutagenesis stuff
Mutagenesis with EMS
Mutagenesis with ENU
Transformation stuff
Yeast transformation
Electrocompetent cells
Heat shock transformation (DH10B homemade)
Protein stuff
BSA conjugation with maleimide (Pierce)
Glycine elution (affinity purification)
IPTG induction
Kinase assay
Cytosolic extract & immunoprecipitation
Cytosolic extract using sonication (fast)
Lee Supplemental
Additional Methods and Materials
GLD-1 mediated translational repression of RME-2 results in yolk uptake only in large oocytes – Photo
Yolk accumulates prematurely in gld-1 (null) (Photo)
UV-crosslink with Probe 9 (5′-end) and Probe 11 (3′-end) of rme-2 mRNA (Photo)
RME-2 expression is regulated normally in fog-2 (null) (Photo)
GLD-1 mediated repression of RME-2 expression is conserved in C. briggsae and C. remanei (Photo)
GLD-1/FLAG is expressed at the appropriate time and place to rescue gld-1 null (Photo)
rme-2 mRNA is expressed at L4 larval stage but RME-2 protein is not expressed (photo)
RME-2 expression is regulated normally in gld-1 (q126/q485) female (Photo)
Schedl Lab
Washington University Genetics
Transformation stuff
Tim Schedl, Ph.D
Department of Genetics
Campus Box #8232
Washington University School of Medicine
4566 Scott Ave.
St. Louis, MO 63110
Contact
PHONE: (314)362-6162
FAX: (314)362-7855