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Requirement for a new transfection assay for Requirement for a new transfection assay for RNAiRNAi
Each cell has unique transfection efficacy.
For successful RNAi, good transfection method is essential.
One easy way is testing with Fluor-conjugated siRNA. As shown in below, the
method is for only transfection but not for RNAi.
Another method is testing gene silencing after transfection using qPCR, which
demands for cost and labor.
A fast and easy transfection assay for RNAi is required.
Fig. 1 Before (left) and after (right) transfection of CEM cell with FITC-conjugated Vimentin siRNA.
Fig. 2 Total RNA sample was prepared after the transfection and used for qPCR for test the silencing of target gene. Unlike HeLa, there was no sign of gene silencing even thought indication of good transfection by the FITC-conjugated siRNA.
Transfection control siRNATransfection control siRNA
Materials included Materials included
• Transfection control siRNA (2 nmole)
• Scramble siRNA (2 nmole)
• siRNA dissolving buffer
The easiest and fastest way for siRNA transfection test
Just add to the cell, wait for 48 hours, and count the viable
cell
The cell toxicity and level of the gene silencing is
proportional
The target gene of the used siRNA is essential for cell
viability
For human, mouse, and rat cells
Transfection control siRNATransfection control siRNA
Cell viability of MCF7 in 48 hour
Scramble siRNA 1 nM control siRNA 1 nM
Gene silencing in 24 hour
*All transfection assay was done by a transfection reagent G-Fectin.
The cell toxicity and level of the gene silencing is proportional
*All transfection assay was done by a transfection reagent G-Fectin.
MCF7
The cytotoxic effect of Transfection control siRNA in mouse and rat cells
*All transfection assay was done by a transfection reagent G-Fectin.
G-Fectin
1. A novel transfection reagent developed for RNAi
2. Whole transfection in 10 min on the same
day. The fastest and easiest method for
transfection.
3. Your gene silencing in 24 hours
4. Lower toxicity
5. Good transfection efficacy
6. Compatible to siRNA. shRNA, and miRNA
7. 0.5ml (for 250 X transfection of 24 well plate)
10 min 10 min transfection transfection
protocol protocol with G-Fectinwith G-Fectin
2 ul of G-FectinsiRNA
Add the mixture to
plate directly
Incubate 24 hrs
qPCR or Western
Add G-Fectin & siRNA to 50 ul of PBS in tube
Incubate 10 min at RT
Detached the cell and add 450 ul of media (5 x 104 cells/
well) to 24 well plate
No cell splitting is required in previous day
G-Fectin has good Transfection efficacy
Transfection assay using Vimentin siRNA final 10 nM
Tested cell line list
The Transfection efficacy was tested using the Transfection Control siRNA and G-Fectin.
• Excellent transfection efficacy with 2ul of G-FectinHeLa, CHO, KB, HT1080, 3T3-L1, COS-1, Caco-2, SKOV3, MDA-MB-468, HEP3B, siHa, MIA-PACA2, PANC1, PLC/PRF/5, MCF7, PC3, C4I, J82, MRC5, HaCat
• Good transfection efficacy with 4 ul of G-FectinSNU-638, SNU-368, SNU-475, Noz, OVCA429, T98G,
• No transfectionJurkat, CEM
G-Fectin has low cytotoxicity
All described cells were spitted in 30 % confluency in 36 hour ahead of viable cell counting with treatment of lipo-plexed scramble RNA in final 10 nM.
G-Fectin KitG-Fectin Kit
Materials included Materials included
• Transfection control siRNA (2 nmole)
• Scramble control siRNA (2 nmole)
• siRNA dissolving buffer
• G-Fectin 0.5ml (for 500 X transfection to 24 well)
G-Fectin + Transfection Control G-Fectin + Transfection Control
siRNAsiRNA
The easiest and fastest way for siRNA
transfection test
Includes all reagents required for
transfection and transfection efficacy
test
Can use to human, mouse, and rat
Recommended