Life Science

RNAi In Situ

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View the entire protocol on BioSupplyNet.com.

View the entire protocol, "In Situ Hybridization of Whole-Mount Mouse Embryos with RNA Probes: Hybridization, Washes, and Histochemistry," at BioSupplynet.com


Thomas Lufkin
This protocol was adapted from "Techniques for Visualizing Gene Products, Cells, Tissues, and Organ Systems," Chapter 16, in Manipulating the Mouse Embryo, 3rd edition, by Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.

ABSTRACT
This is a protocol describing how to perform in situ hybridization on whole mouse embryos. Here we describe the hybridization procedure, and the localization of the DIG-labeled RNA using a conjugate of anti-DIG Fab antibody and calf intestinal alkaline phosphatase. Enzyme activity of the reporter is detected by a color reaction, resulting in the formation of a water-insoluble purple/blue precipitate.
Manipulating the Mouse Embryo - Third Edition
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Products Available for this Protocol

Protocol Material Description Product #  Product Name Add to Cart
Tris-Cl T5941 Trizma® hydrochloride, for molecular biology, Biotechnology Performance Certified, cell culture tested, ≥99.0% (titration) Shopping Cart
NaCl S3014 Sodium chloride, for molecular biology, ≥98% (titration) Shopping Cart
EDTA E5134 Ethylenediaminetetraacetic acid disodium salt dihydrate, for molecular biology, ≥99% Shopping Cart
PBS P5368 Phosphate buffered saline, pH 7.4, Biotechnology Performance Certified Shopping Cart
Proteinase K P6556 Proteinase K from Tritirachium album, lyophilized powder, ≥30 units/mg protein Shopping Cart
MgCl2 M1028 Magnesium chloride solution, for molecular biology, 1.00 M±0.01 M Shopping Cart
Tween 20 P9416 TWEEN® 20, for molecular biology, viscous liquid Shopping Cart
Methanol 34860 Methanol, for HPLC, ≥99.9% Shopping Cart
H2O W4502 Water, Molecular Biology Reagent Shopping Cart
Blocking Reagents
SSC S8015 SSC Buffer Shopping Cart
Formamide F7508 Formamide, for molecular biology, ≥99.5% (GC), liquid Shopping Cart
CHAPS C5070 CHAPS, SigmaUltra, ≥98% (TLC) Shopping Cart
Heparin H4784 Heparin sodium salt, ~170 USP units/mg Shopping Cart
Maleic acid M0375 Maleic acid, ReagentPlus®, ≥99% Shopping Cart
Paraformaldehyde P6148 Paraformaldehyde, reagent grade, crystalline Shopping Cart
KCl P9541 Potassium chloride, for molecular biology, ≥99% Shopping Cart
Na2HPO4 S3264 Sodium phosphate dibasic, for molecular biology, ≥98.5% (titration) Shopping Cart
KH2PO4 P9791 Potassium phosphate monobasic, for molecular biology, ≥98% Shopping Cart
CaCl2 • 2H2O C7902 Calcium chloride dihydrate, cell culture tested, insect cell culture tested, ≥99% Shopping Cart
MgCl2 • 6H2O M2393 Magnesium chloride hexahydrate, cell culture tested, insect cell culture tested Shopping Cart
BM purple AP substrate B3679 BCIP®/NBT-Purple Liquid Substrate System for Membranes Shopping Cart
Sheep serum S2263 Sheep Serum, USA origin, sterile-filtered, cell culture tested Shopping Cart
Sodium azide S8032 Sodium azide, SigmaUltra Shopping Cart

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