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