Qiagens RNeasy kit portal 12-02-2014d1530

2015-02-23

Qiagens RNeasy kit portal 12-02-2014d1530
rna extraction

handbook..manual located here

"C:\Sync\cnio\2014\2014\12 December\12-02-2014d0922\Qiagen RNeasy Mini Handbook 12-02-2014d1525.pdf"

Animal tissue protocol from pg. 36 to 42 of pdf

Qiagens RNeasy kit protocol 12-09-2014d1712

some notes
s: embryo with rneasy kit

didn't really yield any useful results

s: how to isolate rna from mouse embryo

Isolating total RNA from mouse embryos or fetal tissues. (paper)
-http://www.ncbi.nlm.nih.gov/pubmed/21357161
-behind a paywall

s: "mouse embryo" on currentprotocols.com

-Whole Mount In Situ Hybridization and Detection of RNAs in Vertebrate Embryos and Isolated Organs
-http://onlinelibrary.wiley.com/doi/10.1002/0471142727.mb1409s66/pdf

Need to put tissue in RNAlater RNA stabilization reagent or store at -70 C

After disruption and homogenization in Buffer RLT (lysis buffer),
samples can be stored at –70°C for months.

need to disrupt tissue in tissuelyser or tissueRuptor or mortar and pestle

disruption and homogenization are two different things

Cell and tissue lysates can be homogenized using a syringe and needle. Lysate is
passed through a 20-gauge (0.9 mm) needle attached to a sterile plastic syringe at least
5–10 times or until a homogeneous lysate is achieved. Increasing the volume of lysis
buffer may be required to facilitate handling and minimize loss.

homogenization using QIAshredder homogenizer option

Generally, DNase digestion is not required with RNeasy Kits since RNeasy silica-
membrane technology efficiently removes most of the DNA without DNase treatment.

optional on-
column DNase digestion using the RNase-Free DNase Set

Animal tissue protocol from pg. 36 to 42 of pdf