Ordering
Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Plasmid | 50454 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
AAV8 | 50454-AAV8 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart |
This material is available to academics and nonprofits only.
Backbone
- Vector backbonepAAV(Search Vector Database)
- Backbone sizew/o insert(bp)4818
- Total vector size (bp)7937
- Vector typeAAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)Stbl3
- Copy numberLow Copy
Gene/Insert
- Gene/Insert namehM3D(Gq)-IRES-mCitrine
- SpeciesH. sapiens (human)
- Insert Size (bp)3119
- MutationSee supplemental documents for DREADD mutations
- Entrez GeneCHRM3(a.k.a. EGBRS, HM3, PBS)
- Promoterhuman Synapsin 1
- Tag/ Fusion Protein
- HA (N terminal on insert)
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteAsc I(not destroyed)
- 3′ cloning siteNhe I(not destroyed)
- 5′ sequencing primerTCGTGTCGTGCCTGAGAGCG
- 3′ sequencing primerGCATTAAAGCAGCGTATCCACATAGC (Common Sequencing Primers)
Resource Information
- Supplemental Documents
- DREADD information
- Terms and Licenses
- UBMTA
- Ancillary Agreement for Plasmids Containing FP Materials
- genOway Notice of RIghts
- Industry Terms
- Not Available to Industry
- Article Citing this Plasmid
- 1 Reference
Depositor Comments
Please see http://pdspit3.mml.unc.edu/projects/dreadd/wiki/WikiStart for more information.
Information for AAV8 (Catalog # 50454-AAV8)(Back to top)
Purpose
Ready-to-use AAV8 particles produced from pAAV-hSyn-DIO-HA-hM3D(Gq)-IRES-mCitrine (#50454). In addition to the viral particles, you will also receive purified pAAV-hSyn-DIO-HA-hM3D(Gq)-IRES-mCitrine plasmid DNA.
Synapsin-driven, Cre-dependent, excitatory hM3D DREADD expression, with physically separate mCitrine expression for cell body labeling. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 1×10¹³ vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV8 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV8
- PurificationIodixanol gradient ultracentrifugation
- Reporter GenemCitrine (Cre-dependent)
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Ancillary Agreement for Penn Vectors
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
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accctcccgccccccccgtat
(互补碱基不写了)
那么一般来说,你就要导入以accctc结尾的引物使得增幅继续下去,具体方法和注意事项楼主可以看gee_an大侠的资料.
查了一下测序公司的网页,如果要同定微生物,楼主需要准备如下东东:
1、待测菌体.试管或培养皿都可以,但必须是纯粹的待测菌体,不能含有其他的微生物.
2、提纯的DNA.如果1有致病性或传染性,那么楼主就得自己提纯DNA再交给测序公司.DNA纯度要求可以做PCR.
脂代谢及高脂血症的检查
血浆蛋白质检查
诊断酶学
体液平衡紊乱及其检查
R: Reversed primer 反向引物
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