Navigation Links
Housekeeping Genes: Universal Positive,,,Controls in siRNA Knockdown Experiments

RNA interference is a powerful technique for gene knockdown experiments in academic research and an important tool for target identification and target validation in therapeutic research.

A growing number of different cell lines are used for gene knockdown experiments, and in many cases protocols for transfection do not exist. In order to optimize transfection conditions for a new cell line being investigated, luciferase-expressing plasmids are often cotransfected with the siRNA. However, the transfection conditions for plasmids and siRNA are quite different, as the plasmids must enter the nucleus, whereas siRNAs act in the cytoplasm therefore, these optimizations may lead to poor results. In addition, the use of fluorescently labeled siRNAs can give unpredictable results due to hydrophobic effects of fluorophore groups.

We have successfully used housekeeping genes as universal positive control genes for knockdown experiments in two different cell lines. The mRNA levels were analyzed using the LightCycler Instrument and were quantified relative to other housekeeping genes using the LightCycler h-Housekeeping Gene Sets.

Materials and Methods

Cell culture
Cells of the human adherent cell line PC3 were cultured in RPMI 1640 medium containing 10% (v/v) fetal calf serum and 2mM L-glutamine. Cells of the human adherent cell line HCT116 were cultured in McCoys 5A medium containing 10% (v/v) fetal calf serum and 2 mM L-glutamine.

Two days prior to transfection with siRNA oligonucleotides, 5x104 cells were plated in each well of 24- well plates. The siRNA (Dharmac
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Low Abundance cDNA Cloned Using Stratagenes Human Universal cDNA Library
2. Human Universal cDNA Library Array I
3. Universal Guide to DNA Standards
4. Custom and library siRNA for efficient gene silencing
5. Custom and library siRNA for efficient gene silencing
6. Cancer siRNA Oligo Set Version 1.0
7. Library siRNA
8. Custom siRNA Oligo Synthesis Service
9. Efficient RNAi-mediated gene silencing in neuronal cells using QIAGEN siRNA and TransMessenger Transfection Reagent*
10. Quantification of siRNA Silencing Efficiency Using the LightCycler System
11. Confirming gene silencing mechanism by pGFP/GFP22 siRNA co-transfection
Post Your Comments:
(Date:6/2/2015)... Global Stem Cells Group subsidiary Adimarket has been ... for the regeneration and repair of human organs and tissues. ... ambitious expansion plans in Latin America, a commitment made to ... and stem cell therapies. , Bioquark Inc. ... ability to alter the regulatory state of human tissues and ...
(Date:6/2/2015)... June 2, 2015 Meet ARGUS, ... Scanner. Scan & Read People. Analyze emotions and predict ... Concepts, including the very first Responsive-RPG: "The Eye: Genesis". ... , , , , https://www.youtube.com/watch?v=B1ZEtAZ2Yf8 ... truly affect your inner feelings - and those of ...
(Date:6/2/2015)... 02, 2015 “Stem Cell Therapy to help ... and it is used often as an adjunct to surgery ... are not yet aware or comfortable enough with the techniques ... most often reserved for the last resort cases - when ... barely lift a paw to get around. It is an ...
(Date:6/2/2015)... 2015 /PRNewswire/ -- China Cord Blood Corporation (NYSE: CO ) ... leading provider of cord blood collection, laboratory testing, ... announced that the Special Committee of the Company,s board of ... ( China ) Limited as its independent financial ... its United States legal counsel and ...
Breaking Biology Technology:Adimarket Named Latin American Distributor for Bioquark Biologic Products for the Regeneration and Repair of Human Organs and Tissue with Stem Cells 2Adimarket Named Latin American Distributor for Bioquark Biologic Products for the Regeneration and Repair of Human Organs and Tissue with Stem Cells 3ARGUS - The World's First Inner Emotion Scanner Launches on KickStarter 2ARGUS - The World's First Inner Emotion Scanner Launches on KickStarter 3VetStem Biopharma, Proudly Announces a New Long Term Partnership with VCA West Los Angeles in Support of Stem Cell Therapy Training and Clinical Opportunities. 2VetStem Biopharma, Proudly Announces a New Long Term Partnership with VCA West Los Angeles in Support of Stem Cell Therapy Training and Clinical Opportunities. 3China Cord Blood Corporation's Special Committee Appoints Independent Financial Advisor and Legal Counsel to Evaluate Non-binding "Going Private" Proposal 2China Cord Blood Corporation's Special Committee Appoints Independent Financial Advisor and Legal Counsel to Evaluate Non-binding "Going Private" Proposal 3
... Inc. (OTC Bulletin Board: GOVX), an Atlanta-based biotechnology company ... its financial results for the fourth quarter and year ... recent operational highlights for calendar year 2009 to date.GeoVax ... quarter ended December 31, 2008, as compared to a ...
... Micromet, Inc. (Nasdaq: MITI ), a biopharmaceutical ... cancer, inflammation and autoimmune diseases, today announced that it ... North America to its most advanced BiTE antibody candidate, ... conducting a phase 2 clinical trial with blinatumomab for ...
... Production Efficiency and Reliability for New Shire Human Genetic ... HON ) today announced it will help ... development capabilities for creating life-saving medications. Shire Human Genetic ... will use several Honeywell solutions - including Experion(R) ...
Cached Biology Technology:GeoVax Labs, Inc. Reports Fourth Quarter and Year-End 2008 Financial Results 2GeoVax Labs, Inc. Reports Fourth Quarter and Year-End 2008 Financial Results 3GeoVax Labs, Inc. Reports Fourth Quarter and Year-End 2008 Financial Results 4GeoVax Labs, Inc. Reports Fourth Quarter and Year-End 2008 Financial Results 5GeoVax Labs, Inc. Reports Fourth Quarter and Year-End 2008 Financial Results 6GeoVax Labs, Inc. Reports Fourth Quarter and Year-End 2008 Financial Results 7GeoVax Labs, Inc. Reports Fourth Quarter and Year-End 2008 Financial Results 8GeoVax Labs, Inc. Reports Fourth Quarter and Year-End 2008 Financial Results 9GeoVax Labs, Inc. Reports Fourth Quarter and Year-End 2008 Financial Results 10GeoVax Labs, Inc. Reports Fourth Quarter and Year-End 2008 Financial Results 11Micromet, Inc. To Regain Rights to Blinatumomab in North America; Reports Financial Results for the Fourth Quarter and Full Year ended December 31, 2008 2Micromet, Inc. To Regain Rights to Blinatumomab in North America; Reports Financial Results for the Fourth Quarter and Full Year ended December 31, 2008 3Micromet, Inc. To Regain Rights to Blinatumomab in North America; Reports Financial Results for the Fourth Quarter and Full Year ended December 31, 2008 4Micromet, Inc. To Regain Rights to Blinatumomab in North America; Reports Financial Results for the Fourth Quarter and Full Year ended December 31, 2008 5Micromet, Inc. To Regain Rights to Blinatumomab in North America; Reports Financial Results for the Fourth Quarter and Full Year ended December 31, 2008 6Micromet, Inc. To Regain Rights to Blinatumomab in North America; Reports Financial Results for the Fourth Quarter and Full Year ended December 31, 2008 7Micromet, Inc. To Regain Rights to Blinatumomab in North America; Reports Financial Results for the Fourth Quarter and Full Year ended December 31, 2008 8Micromet, Inc. To Regain Rights to Blinatumomab in North America; Reports Financial Results for the Fourth Quarter and Full Year ended December 31, 2008 9Micromet, Inc. To Regain Rights to Blinatumomab in North America; Reports Financial Results for the Fourth Quarter and Full Year ended December 31, 2008 10Micromet, Inc. To Regain Rights to Blinatumomab in North America; Reports Financial Results for the Fourth Quarter and Full Year ended December 31, 2008 11Micromet, Inc. To Regain Rights to Blinatumomab in North America; Reports Financial Results for the Fourth Quarter and Full Year ended December 31, 2008 12Honeywell Technology Selected to Control Manufacturing Processes at Major U.S. Biotech Site 2Honeywell Technology Selected to Control Manufacturing Processes at Major U.S. Biotech Site 3Honeywell Technology Selected to Control Manufacturing Processes at Major U.S. Biotech Site 4
(Date:5/11/2015)... Curemark LLC, a privately held drug research ... Phase III double blind, randomized, placebo-controlled clinical trial to ... all children ages 3-8 with Autism. Previously, Curemark announced ... blinded clinical trial for CM-AT in children ages 3-8 ... enzyme chymotrypsin. This new trial will help determine whether ...
(Date:5/7/2015)... Sweden , May 7, 2015 ... touch fingerprint sensors, FPC1022 and FPC1035, FPC,s smallest ... and FPC1035 are mainly considered for integration on ... size gives smartphone OEMs increased possibilities to integrate ... The decreased size also improves possibilities for module ...
(Date:4/27/2015)... more than four decades, AUVSI,s Unmanned Systems ... event for the unmanned land, sea, and air industries. ... local, national, or trade news organization interested in gathering ... applications of unmanned technologies. The conference will explore the ... and how it will soon impact the lives of ...
Breaking Biology News(10 mins):Curemark, LLC, Launches New Phase III Trial in Expanded Population of Children with Autism 2FPC Introduces its Smallest Touch Fingerprint Sensors to Date 2UNMANNED SYSTEMS 2015: Largest International Conference and Trade Show for the Drone and Unmanned Technology Industry 2UNMANNED SYSTEMS 2015: Largest International Conference and Trade Show for the Drone and Unmanned Technology Industry 3UNMANNED SYSTEMS 2015: Largest International Conference and Trade Show for the Drone and Unmanned Technology Industry 4UNMANNED SYSTEMS 2015: Largest International Conference and Trade Show for the Drone and Unmanned Technology Industry 5
... June 2009 Elsevier, the leading publisher of scientific, ... the highlights of its journal impact factor performance in ... factors (IF) increase from 2007 to 2008. According to ... journals took the #1 position in 51 categories (of ...
... 2008 Olympic Games, world record times of the men,s 100m ... and technological advances. Imagine if this improvement could be achieved ... Research Institute of Wildlife Ecology in Austria have managed to ... high in polyunsaturated fatty acids. Polyunsaturated fatty acids are ...
... a changing climate depends on the nutritional value and yield ... Australia have found an increase in toxic compounds, a decrease ... under high CO 2 and drought conditions. ... 29 June 2009 at the Society for Experimental Biology Annual ...
Cached Biology News:Elsevier announces 2008 journal impact factor highlights 2Elsevier announces 2008 journal impact factor highlights 3Mice run faster on high-grade oil 2
...
... Immunoperoxidase Secondary Detection System uses biotin-avidin-HRP complexed ... in the Primary Antibody. The primary antibody ... specimen. Any excess antibody is removed by ... to biotin) is added and will react ...
... Designed specifically for high ... cells. Low toxicity reagent proven ... wide range of cell types. ... than 95% of target gene ...
... Powerful exact mass measurements in ... orthogonal electrospray time-of-flight mass spectrometer (ESI-oa-TOF) ... machine in a format, which fits ... a mass resolution exceeding 10,000 FWHM ...
Biology Products: