0610009B22Rik AAV

10006101 μg, 5 x 200 μl


DescriptionThis 0610009B22Rik ready-to-use AAV is part of abm’s AAV Expression System and can be used directly to transiently over-express the 0610009B22Rik gene in a wide range of host cells or animal models.
Accession NumberNM_025319
Unit quantity1 μg, 5 x 200 μl
Titer>1 x 109 GC/ml
Gene Name0610009B22Rik
SpeciesThis gene is available from: Mouse
Insert SizeNM_025319:423
Vector SizepAAV-G-CMV:4687
Related SequencesNM_025319:NM_025319.2,GI:141801968
Shipping ConditionsAAV vectors are shipped with gel ice packs. AAV viruses are shipped with dry ice.
Storage ConditionStore AAV vectors at -20°C. Store AAV viruses at -80°C. Avoid repeated freeze-thaw cycles by storing in small aliquots.
Guaranteeabm guarantees that the correct ORF construct is provided and the mRNA expression is displayed upon successful transduction. If this is not the case, we will provide a one-time replacement. Customers must provide adequate data to show >80% transfection efficiency with a positive control, plus additional qPCR data or a western blot to evaluate the level of gene expression. The replacement will not be covered by the same guarantee.
Please note that due to the large number of variables applicable, any further expression analysis (e.g. protein expression) is not covered by the guarantee, as such analysis is dependent on the end user's experimental conditions.
Disclaimer1) 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.
Accession NumberNM_025319

What is the difference between Retro-, Lenti-, and Adeno- viruses?
Retrovirus: Classic, can integrate into the genome but with low transduction efficiency. They are useful for gene transfer and protein expression in cells that have low transfection efficiency with other transfection reagents. Lentivirus: Can integrate into the genome with relatively high transduction efficiency and they are very useful for cells that have low transfection efficiency with other transfection reagents. No special competent cells required, as they are stable plasmids. Lentiviruses are a powerful tool for stable gene transfer to both dividing and non-dividing cells in vitro and in vivo. Adenovirus: Only work transiently (about 7 days) but have almost 100% transduction efficiency. Adenoviruses can infect a broad range of cell types with the highest efficiency and infection is not dependent on active host cell division. A second key feature is that high virus titers and high-level gene expression can be obtained in most mammalian cells.
What are the correct concentration units for each recombinant viral particle?
For lentiviruses and retroviruses, they are measured in CFU/ml (colony-forming units per millilitre). Transduction with lentiviruses and retroviruses can cause the formation of colonies, which can be quantified for concentration. For AAV the titer is measured as genome copies per mL (GC/mL). Adenoviruses are measured as PFU/ml (plaque-forming units per millilitre). Transduction with adenoviruses will kill packaging cells, forming plaques in the process for quantification. The concentration for all three types of viruses can also be classified as IU/ml (Infectious Units/ml). Ultimately, the units refers to the viral particles and different units reflect the different assays involved.
How long after transduction can the infection efficiency be observed?
You can observe transduction efficiency from 48 hours up to 5 days after infection.
How much DNA do I need to provide for Custom AAV without DNA Amplification?
You will need to provide purified plasmid DNA at a concentration of 0.5ug/ul or more.
50 ug DNA needed for Custom AAV without DNA amplification (10^9 GC/ml)
200 ug DNA needed for Custom AAV without DNA amplification (10^12 GC/ml)
300 ug DNA needed for Custom AAV without DNA amplification (10^13 GC/ml)
What's the difference between GC/ml and vg/ml?
Both are used interchangeably and it is a qPCR based titer method.
Why do I not see GFP expression after infecting AAV in my cell line?
Serotype selection is an important parameter affecting transduction ability of AAV particles. Thus, determine which serotype works best for your cell line. You could refer to our technical sheet "AAV - General Guideline to Serotype Selection". eg. Serotype 5 has limited transduction ability on most cell types.
Is your AAV preparation in-vivo grade?
Our high titer AAV preps undergo extensive purification steps leading to high quality viral particles ready to inject for your in-vivo models. For reference, see our in-vivo infectivity data as tested by an independent lab {https://www.abmgood.com/AAV-Adeno-Associated-Virus.html}
What are the QC methods for testing AAV?
We provide qPCR based titer as primary method of determining successful packaging. If your virus has GFP or RFP reporter, we also perform virus infectivity testing in HEK293T cell line and provide you image in CoA.Since infectivity is serotype dependent, and HEK293T cells are not infected well with Serotype 4,5 and 6, we are unable to confirm infection for these serotypes in our standard evaluation cell line. For such cases, titer as determined by qPCR is considered final parameter to determine successful packaging of virus.
Can I work with different serotypes of AAV virus in the same equipment to keep the infected cells?
There is no problem with using different serotypes in the same equipment, as long as the handler takes the basic precautions to avoid cross-contamination.

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