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Selected Protocol from "RNAi, A Guide to Gene Silencing", Gregory J. Hannon.
Mammalian RNA Interference, Thomas Tuschl (Chapter 13, pg. 280-288)
Microinjection of dsRNA into Mouse Oocytes and Early Embryos
ABSTRACT
This protocol describes how to introduce a double-stranded RNA (dsRNA) of choice into mouse oocytes or fertilized one-cell embryos by microinjection. For collection of mouse oocytes and early embryos, see Collection of Mouse Oocytes for RNAi and Collection of Early Mouse Embryos for RNAi. It is advisable to confirm knock-down of the gene of interest by RT-PCR (see RNA Isolation and RT-PCR from dsRNA-treated Mouse Oocytes and Early Embryos). |
Buy this book, Z701130
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Products Available for this Protocol
| Protocol Material Description |
Product # |
Product Name |
Add to Cart |
| Buffers,
Solutions, and Reagents |
|
|
|
| NaCl |
S3014 |
Sodium chloride |
|
| KCl |
P9541 |
Potassium chloride |
|
| K2HPO4 |
P9666 |
Potassium phosphate dibasic trihydrate |
|
| MgSO4 |
M2643 |
Potassium phosphate dibasic trihydrate |
|
| CaCl2 |
21115 |
Calcium chloride solution |
|
| NaHCO3 |
S3817 |
Sodium bicarbonate, Biotechnology Performance Certified, cell culture tested |
|
| Sodium lactate |
L4263 |
Sodium DL-lactate solution |
|
| Sodium |
483745 |
Sodium |
|
| DMSO |
D8418 |
Dimethyl sulfoxide for molecular biology |
|
| Streptomycin |
S9137 |
Streptomycin sulfate salt |
|
| L-glutamine |
G6392 |
L-Glutamine cell culture tested, γ-irradiated |
|
| EDTA |
E7889 |
Ethylenediaminetetraacetic acid disodium salt solution |
|
| Bovine serum albumin (BSA) |
B6917 |
Albumin from bovine serum, for molecular biology, Non-acetylated, lyophilized powder |
|
| Gentamicin |
G1272 |
Gentamicin solution |
|
| Phenol Red |
P3532 |
Phenol Red, powder, cell culture tested |
|
| Polyvinylpyrrolidone (PVP) |
P5288 |
Polyvinylpyrrolidone, for molecular biology, nucleic acid hybridization tested |
|
| Glucose |
G5400 |
D-(+)-Glucose |
|
| Penicillin G |
P3032 |
Penicillin G sodium salt |
|
| IBMX (3-isobutyl-1-methylxanthine) |
I7018 |
3-Isobutyl-1-methylxanthine, SigmaUltra |
|
| HEPES (pH 7.2) |
51558 |
HEPES buffered saline, for molecular biology, 2× concentrate |
|
| Sodium pyruvate |
P5280 |
Sodium pyruvate, powder, cell culture tested, insect cell culture tested |
|
| Bicarbonate-free minimal essential medium (Earle's salts) |
E7510 |
10 ×, Without sodium bicarbonate, sterile-filtered, liquid, cell culture tested |
|
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