Inhibitor hsa-let-7a-2-3p AAV miRNA Virus (Serotype 3)
|Amh3000303||5 x 200 μl|
|Description||This ready-to-use AAV is part of abm’s AAV Expression System and can be used directly to knockdown your miRNA of interest in a wide range of host cells or animal models.|
|Species||Human (H. sapiens)|
|Titer||>1 x 109 GC/ml|
|Unit quantity||5 x 200 μl|
|Serotype||AAV Serotype 3|
|Storage Condition||AAV viruses are shipped with dry ice. For long term storage, it is recommended to store the viruses at -80°C in small aliquots to avoid repeated freeze-thaw cycles.|
|QC||qPCR Based Titer Determination and Transduction Efficiency Test|
|Recommended Controls||pAAV-mir-OFF-hGH-amp-Blank Control Vector(Cat# Am00700a)|
|Mammalian Selection Marker||Puromycin|
|Disclaimer||1) Disclaimer for Transcript Variants: The provided accession number refers to the transcript (mRNA) sequence for this product. The molecular sequence of this clone aligns with the gene accession number as a point of reference only. However, individual transcript sequences of the same gene can differ through naturally occurring variations (e.g. polymorphisms), each with its own valid existence. This clone is substantially in agreement with the reference, but a complete review of all prevailing variants is recommended prior to use. All sales are final.|
2) Disclaimer for Gene Sequence: The provided accession number refers to the transcript (mRNA) sequence for this product. Please verify that this is the desired transcript sequence by cross-referencing. This is important because a single gene can have multiple different transcripts owing to naturally occurring variations. All sales are final.
3) Disclaimer for Intended Use: All of abm's vectors and viral particles are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its vector(s) in therapeutic/diagnostic application(s).
4) Disclaimer for Extra Nucleotides: Cloning may lead to the insertion of extra nucleotides at the 5' or 3' end of the target sequence which, in most cases, is innocuous to the stability/functionality of the construct.
5) abm guarantees that at least 1 out of the 3 sgRNA constructs purchased in a set designed to be used with Cas9 Nuclease will result in gene knock-out due to frameshift mutations in over 50% of cells, after successful infection and drug selection. This guarantee applies to sgRNAs designed to target human, mouse or rat genes only. If knock-out is not achieved in extremely rare cases, a one-time replacement of another set of 3 targets with alternative sgRNA sequences will be provided. To qualify for this replacement, customers must examine knock-out efficiency by Surveyor assay. Before sending your inquiry, please make sure you have optimized your experiments as far as possible. This includes (where applicable) increasing and optimizing your MOI, increasing the duration of infection (up to 72 h), and carrying out clone screening before assaying for knock-out. Please also provide data to show that a reporter virus was used to optimize the MOI for your target cell line. Customers must provide adequate data to show >80% infection efficiency with a positive control, plus additional qPCR data to evaluate the level of mRNA expression.
For vector transfection, please evaluate the vector transfection efficiency by detecting Cas9 or puromycin expression for the 'All-in-One' vectors using qPCR, or neomycin for constructs containing only the sgRNA. In addition, please provide Surveyor Assay or Sanger Sequencing data on at least 20 isolated clones.
abm limits its obligation and liability for the success of this technology to providing one replacement of any sgRNA lentivector product only. The replacement set will not be covered by the same guarantee. If these constructs are also considered to be ineffective then the gene is most likely not susceptible to sgRNA knock-out.
|What is the Sanger miRBase Sequence Database?|
miRBase is a sequence database that has been established by the Sanger Institute. Each entry in the microRNA Registry represents a predicted hairpin portion of a microRNA transcript (termed mir in the database), with information on the location and sequence of the mature microRNA sequence (termed miR). The database provides microRNA gene hunters with unique names for novel microRNA genes prior to publication of results and a searchable database of published microRNA.
|When was the latest update of your array sequences?|
The content of our arrays has been updated to miRBase Release 16.0.
|Do you sell any products that allow detection of microRNAs using in-situ methods|
We currently don't have in-situ miRNA detection methods.
|Do you have any mutant constructs available for the 3'UTR miRNA reporter vectors for miRNA-mRNA interaction validation?|
We do not have any mutant constructs in our inventory, all the listed constructs are wild type. We can offer mutant constructs as a custom service. The exact sequence to be mutated must be supplied by the end-user in each case. Please contact [email protected] with any specific requests.
|Do I need to use an internal control primer for quantitative detection of miRNAs (the same as housekeeping primers which we use in qPCR for mRNAs detection)?|
For human miRNA plates, the controls are as follows: 1) SNORD44, cat MPH00003 2) SNORD47, cat MPH00004 3) SNORD48, cat MPH00005 4) U6-2, cat MPH00001
|How does miRNA inhibitor adenovirus work?|
The miRNA inhibitor is in the form of a short hairpin which is complementary to the mature miRNA sequence that it targets. Its expression is under the control of the H1 promoter in an Adenovirus vector. The packaged adenovirus can be used to transfect your target cells and deliver the miRNA inhibitor into the cells and knockdown its target mature miRNA.
|Can I quantify mature miRNA in total RNA using your cDNA synthesis and kit and qPCR master mix?|
Yes, that is what it is designed for.
|Which genes are targeted by a specific miRNA?|
You can search for predicted and validated miRNA target genes at http://mirbase.org/. Just type in your miRNA name (eg. hsa-mir-145) or accession number in the search bar and look for the targeted genes.
|Does the profiling (qPCR) require a specific instrument or it can be done by every real time PCR instrument?|
We have a range of miRNA qPCR mastermixes available in optimized formulations for all qPCR machines, please see our web page http://www.abmgood.com/miRNA-qPCR-Mastermixes-microRNA-qPCR.html
|pLenti-III-miR-Off system or LentimiRa system uses blank vector for control, but I usually use scrambled control for si-RNA experiments. Is it supported to use blank control not scrambled for miRNA experiments?|
Both scrambled and blank controls are effective negative controls. However, there are debates over off-target effects when using scrambled sequences. By using a blank vector as negative control, the off-target effect can be eliminated. If your experiment requires a scrambled control over a blank control, we can custom make that for you as well upon request.
|Why are miRNA mimics on your site offered at 2 x 2.5nmol, but miRNA inhibitors at 2 x 5.0nmol?|
This is because our mimics now are double-stranded while the inhibitors are single-stranded. The amount of materials are quantified in OD and both mimics/inhibitors are synthesized to yield the same OD. Therefore, the number of moles of mimics is half of that of the inhibitors.
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