Navigation Links
Selecting siRNA Sequences to Incorporate into the pSilencer Vectors


The first step in preparing a plasmid for siRNA experiments is to identify target sequences in the gene of interest that are susceptible to siRNA-induced degradation. We have found that a little more than half of the siRNAs provide at least a 50% reduction in target mRNA levels and approximately 1 out of 4 siRNAs provide a 75-95% reduction. The general process begins with scanning the length of the target gene for potential siRNA target sites. For siRNA expression vectors, the target sites should have 5' terminal AAs because, upon folding the siRNAs hairpins will give rise to UU overhangs. These UU overhangs will therefore be complementary to the AA in the target site. The 21 nucleotide siRNA target sequences are then compared to an appropriate genome database to eliminate sequences with significant homology to other genes. For screening, we typically test four siRNAs per target. We space the siRNAs down the length of the gene sequence to reduce the chances of targeting a region of the mRNA that is either highly structured or bound by regulatory proteins.

Constructing four siRNA expression plasmids for each target can be time-consuming and expensive. In vitro transcription provides a less formidable siRNA screening method. To ensure that siRNAs expressed from plasmids are functionally equivalent to siRNAs prepared by in vitro transcription, we prepared plasmids and siRNAs targeting four different sequences in the cyclophilin and GAPDH genes. These nucleic acids were transfected into HeLa cells. Silencing was evaluated by Northern analysis using probes specific to GAPDH, cyclophilin, and 28S rRNA. The hybridiz
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Custom and library siRNA for efficient gene silencing
2. Custom and library siRNA for efficient gene silencing
3. Cancer siRNA Oligo Set Version 1.0
4. Library siRNA
5. Custom siRNA Oligo Synthesis Service
6. Efficient RNAi-mediated gene silencing in neuronal cells using QIAGEN siRNA and TransMessenger Transfection Reagent*
7. Quantification of siRNA Silencing Efficiency Using the LightCycler System
8. Housekeeping Genes: Universal Positive Controls in siRNA Knockdown Experiments
9. Confirming gene silencing mechanism by pGFP/GFP22 siRNA co-transfection
10. siRNA transfection optimization with the Agilent 2100 bioanalyzer
11. siRNA Design Guidelines
Post Your Comments:
(Date:6/29/2015)... , June 29, 2015 BioNano Genomics, ... of top research institutes, announced today that a collaboration ... Sinai resulted in the creation of the first comprehensive ... single DNA molecule methods for sequencing and genome mapping. ... provided the comprehensive analysis of the contiguous diploid human ...
(Date:6/29/2015)... June 29, 2015  Neuralstem, Inc. (NYSE MKT: CUR), ... develop small molecule and cell therapy treatments for central ... approved for listing on the NASDAQ Capital Market under ... to trade on the NYSE MKT until the market ... Capital Market is expected to commence on July 13, ...
(Date:6/29/2015)... June 29, 2015   Veracyte, Inc . (Nasdaq: ... field of molecular cytology, today announced that it has ... Top Workplaces list for the second year in a ... solely on an annual survey of Bay Area-company employees. ... the degree to which Veracyte values our employees and ...
(Date:6/26/2015)... , June 26, 2015  SeraCare Life ... diagnostics manufacturers, today announced that it has signed ... the National Cancer Institute (NCI), part of the ... and positive controls for cancer assays.  The CRADA ... and accurately quantitate control DNA biosynthetics spiked into ...
Breaking Biology Technology:BioNano Mapping Technology Achieves Most Contiguous Clone-free Human Genome Assembly to Date 2BioNano Mapping Technology Achieves Most Contiguous Clone-free Human Genome Assembly to Date 3Neuralstem, Inc. Announces Approval for Listing on the NASDAQ Capital Market 2Neuralstem, Inc. Announces Approval for Listing on the NASDAQ Capital Market 3Veracyte Named a 2015 "Top Workplace" in the San Francisco Bay Area 2Veracyte Named a 2015 "Top Workplace" in the San Francisco Bay Area 3SeraCare Life Sciences Signs CRADA with NCI to Advance Cancer Diagnostics 2SeraCare Life Sciences Signs CRADA with NCI to Advance Cancer Diagnostics 3
... PRINCETON, New Jersey and SAN DIEGO,California, September 8 , ... for Continuous Process,Chemistry Technologies and for Further Global Expansion, ... investment of $3.3M,by a group of private and financial ... Graphisoft and Dave Tapolczay, CEO of Medical,Research Council Technology ...
... announced that it has appointed A.J. Whalen as Solutions Director. ... ... 8, 2008 -- www.symphony-consulting.com [Symphony Management Consulting] announced ... , , ,Mr. Whalen joins Symphony from Wellesley Information Services ...
... Cell Therapeutics, Inc.,(CTI) (Nasdaq: CTICD ; ... a letter from the Listing Qualifications Staff of ... is not in compliance with the,$50,000,000 minimum market ... Marketplace Rule 4450(b)(1)(A). Furthermore, the Company does not,comply ...
Cached Biology Technology:World Leader in Bench-Top Flow Chemistry Instruments ThalesNano, Inc. Closes Funding Round 2Symphony Managemnt Consulting Hires New Solutions Director 2CTI Receives NASDAQ Letter 2CTI Receives NASDAQ Letter 3
(Date:6/25/2015)... , June 25, 2015  TAKE Solutions Ltd., ... patent by the United States Patent and Trademark Office ... This process leverages TAKE Solutions, Clinical Accelerators to reduce ... 50% (when compared to standardization without the accelerators), thus ... At the heart of the ...
(Date:6/24/2015)... , June 24, 2015 Biometry authentication ... emerging biometric password solution one face in. ... one face in, entering the expanding ... and affordable multifactor-authentication biometric answer to the password ... provided in partnership with KeyLemon, one face in ...
(Date:6/18/2015)... NEW YORK , June 18, 2015 ... in US$ Million by the following Product Segments, which ... Immunoassay Systems, Chemiluminescence Immunoassay Systems, Radio Immunoassay Systems, and ... for the US, Canada , ... Asia-Pacific , Latin America ...
Breaking Biology News(10 mins):TAKE Solutions Awarded Patent By USPTO 2KeyLemon Announces Partnership With Biometric Password Solution onefacein 2Global Immunoassay Systems Market 2Global Immunoassay Systems Market 3Global Immunoassay Systems Market 4Global Immunoassay Systems Market 5Global Immunoassay Systems Market 6Global Immunoassay Systems Market 7Global Immunoassay Systems Market 8Global Immunoassay Systems Market 9Global Immunoassay Systems Market 10Global Immunoassay Systems Market 11Global Immunoassay Systems Market 12Global Immunoassay Systems Market 13Global Immunoassay Systems Market 14Global Immunoassay Systems Market 15Global Immunoassay Systems Market 16Global Immunoassay Systems Market 17Global Immunoassay Systems Market 18Global Immunoassay Systems Market 19Global Immunoassay Systems Market 20Global Immunoassay Systems Market 21Global Immunoassay Systems Market 22Global Immunoassay Systems Market 23Global Immunoassay Systems Market 24Global Immunoassay Systems Market 25Global Immunoassay Systems Market 26Global Immunoassay Systems Market 27Global Immunoassay Systems Market 28Global Immunoassay Systems Market 29
... termites use vibrations to choose the wood they eat ... in homes and also may provide insights into the ... most noise but have some signals that trigger attention ... entomologist Theo Evans says laboratory experiments have found that ...
... A team of Fox Chase Cancer Center scientists led ... genetic mutation that keeps a mouse strain from developing ... The report by Kappes and his colleagues appears in ... first discovered the mice with this naturally occurring defect ...
... 2,or ACC2, appears to turn the adipose or fat ... the animals can eat more and weigh less,than their ... appears online today in the Proceedings of the National,Academy ... mice bred to lack ACC2," said Dr. Salih,Wakil, chair ...
Cached Biology News:Termites feed through good vibrations 2Fox Chase Cancer Center scientists identify immune-system mutation 2Lack of enzyme turns fat cells into fat burners 2
Leptin Receptor Polyclonal Ab 100 ug purified rabbit anti-mouse Leptin Receptor. Tested in Western blotting....
... AccuPrime™ Pfx DNA Polymerase is ideal ... for downstream applications such as cloning ... provided by a proprietary enzyme preparation ... sp. with proofreading (3'Cut Site5' exonuclease) ...
... 177 amino acid polypeptide sequence corresponding to ... Produced by activation of a recombinant ... coli periplasm. SPECIFIC ACTIVITY:The specific activity ... mU/mg, where 1 U is the activity ...
... is a highly purified preparation qualified to ... vectors. The preparation is functionally tested for ... of detectable contaminating proteases. Prepared from human ... tests to be negative for HBsAg and ...
Biology Products: