followed throughout the procedure.
Proper microbiological, aseptic technique should always be used when working
with RNA. Hands and dust particles may carry bacteria and molds, and are
the most common sources of RNase contamination. Always wear latex or vinyl
gloves while handling reagents and RNA samples to prevent RNase contamination
from the surface of the skin. Change gloves frequently and keep tubes closed.
Ensure that all glassware, plasticware, and solutions (including water and
culture medium) are RNase-free.
The use of sterile, disposable, plastic tubes throughout the procedure is
recommended. These tubes are generally RNase-free and do not require pretreatment
to inactivate RNases.
No currently available purification method can guarantee that RNA is completely
free of DNA, even when no DNA is visible on an agarose gel. For RNA transfection,
digestion of the purified RNA with RNase-free DNase is recommended. The
RNA can then be repurified using the RNeasy Kit.
Optimizing RNA Transfection
To achieve optimal transfection efficiency for every cell typeRNA
combination, we recommend optimizing a number of parameters:
The amounts of TransMessenger Transfection Reagent, RNA, and TransMessengerRNA
The cell number/confluency
The length of exposure of cells to TransMessengerRNA complexes
Once the parameters yielding maximum transfection efficiency have been determined,
they should be kept constant in every experiment usi
Source:Page: All 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 Related biology technology :1
. TransMessenger Transfection Reagent2
. Efficient DNA transfection of primary CNS neurons using TransMessenger
. TransMessenger Transfection Reagent4
. Efficient RNAi-mediated gene silencing in neuronal cells using QIAGEN
siRNA and TransMessenger Transfection Reagent*5
. Highest Transfection Efficiency of an Endotoxin-Sensitive Mammalian Cell
. Optimizing Transfection Conditions for Studying Signal Transduction Pathways7
. Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells8
. Improve Lipid- or Calcium Phosphate-Mediated Transfection of Human Dermal
. Versatile Transfection Reagent Offers Low Toxicity and Consistent
. Low-Toxicity, Lipid-Mediated Transfection of Mammalian Cells11
. Mycoplasma Contamination Reduces the Effect of Lipid-Mediated Transfection
of Mammalian Cells