Navigation Links
RNAi: Size Does Matter


Everyone wants to design the dsRNA that most efficiently silences the gene of interest. But how does dsRNA length correlate with silencing efficiency? Ambion scientists addressed this question in the
following experiment.

dsRNA of different sizes and directed against different regions of hrp48 and U2AF50 cDNA (Figure 1A), were prepared using the MEGAscript RNAi Kit and tested on Drosophila L2 cells. For both targeted genes, Western blot analyses showed a stronger silencing effect with dsRNA corresponding to the full length cDNA than with shorter dsRNAs (Figure 1B and D). Similar reduced activities (in comparison to that of full length dsRNAs) were obtained with shorter dsRNA specific for either the 5' or the 3' end of U2AF50 cDNA (Figure 1B).

Interestingly, the high level of silencing induced by the longer dsRNAs could be reached with the shorter dsRNAs by increasing the concentration of the shorter dsRNAs 2-3 fold (Figure 1D). Because yields of dsRNA synthesized and purified with the MEGAscript RNAi Kit are reproducibly higher for shorter dsRNA (Figure 1C), two to five time more experiments can be performed per transcription reaction with 300 to 600 nt long dsRNA. Therefore, it may be more convenient and cost effective to perform silencing experiments with the higher concentration of shorter dsRNA.

Figure 1. Correlation Between dsRNA Size, Transcription Yield and Silencing Effect. (A) Schematic representation of the various dsRNA used in this study. (B) Western blot ana lysis 72 hours post treatment with 10 nM of the indicated dsRNA. (C) Yields of dsRNA synthesized and purified with the MEGAscript RNAi kit using 0.4 g of PCR DNA templates containing T7 RNA polymerase promoters on both ends. (D) Western blot analysis 72 hr post treatment with different concentrations of the indicated dsRNA.


back to top


Ordering Information
Cat# Product Name Size 1626 MEGAscript RNAi Kit 20 rxns
'"/>

Source:


Page: All 1 2

Related biology technology :

1. RNAi: A How To for New Users
2. RNAi: Get the Whole Story
3. RNAi: The Controls You Need
4. siRNA screening of the cell cycle with two dynamic GFP sensors (from Discovery Matters Issue 1, July 2005)
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:1/21/2017)... Colo. , Jan. 20, 2017 ... or the "Company"), announced that on January 14, 2017 ... plan under which the Company will terminate certain employees ... Bioptix Diagnostics, Inc.  The Company commenced terminations on January ... within 30 days.  The Company may pay severance benefits ...
(Date:1/21/2017)... , Jan. 21, 2017   Boston Biomedical , ... designed to target cancer stemness pathways, today presented data ... napabucasin, at the 2017 American Society of Clinical Oncology ... . In a Phase Ib/II ... designed to inhibit cancer stemness pathways by targeting STAT3 ...
(Date:1/20/2017)... , January 20, 2017 Stock-Callers.com ... conditions have influenced the most recent performances of select ... (NASDAQ: RGLS ), Abeona Therapeutics Inc. (NASDAQ: ... TBPH ), and Sage Therapeutics Inc. (NASDAQ: ... by Grand View Research, global Biotech market size is expected to ...
(Date:1/20/2017)... NEW YORK , January 20, 2017 ... Health Organization, cancer is one of leading causes of ... in 2012. Although the number of cancer related deaths ... since 1990. Rising in incidence rate of various cancers ... According to a research report by Global Market Insights, ...
Breaking Biology Technology:
(Date:1/19/2017)... PUNE, India , January 19, 2017 According to ... Market, Opportunities and Forecast, 2014 - 2022," the global biometric sensor market is ... from 2016 to 2022. In 2015, Asia-Pacific dominated the ... public and private sectors. Continue Reading ... ...
(Date:1/18/2017)... Md. , Jan. 18, 2017  In vitro ... respect to mergers and acquisitions (M&A), and Kalorama Information ... for such acquisitions have been shifting. Generally, uncertainty in ... and the U.S. has changed the acquisitions landscape. ... has resulted in companies buying partners outside of their ...
(Date:1/13/2017)... 13, 2017 Sandata Technologies, LLC, a ... homecare industry, including Electronic Visit Verification™ (EVV™), announced ... Jugs, as Senior Vice President of Product Management. ... of homecare experience to Sandata, where he will ... to align Sandata,s suite of solutions with the ...
Breaking Biology News(10 mins):