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Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Plasmid | 104052 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
AAV PHP.V1 | 104052-AAVPHP.V1 | Virus (100 µL at titer ≥ 7×10¹² 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)5456
- Vector typeAAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)NEB Stable
- Copy numberHigh Copy
Gene/Insert
- Gene/Insert nameEYFP
- Insert Size (bp)720
- PromoterCAG
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteAscI(not destroyed)
- 3′ cloning siteNheI(not destroyed)
Resource 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 visitimages/Addgene/246405v2 for bioRxiv preprint.
Information for AAV PHP.V1 (Catalog # 104052-AAVPHP.V1)(Back to top)
Purpose
Ready-to-use AAV PHP.V1 particles produced from pAAV-CAG-DIO-EYFP (#104052). In addition to the viral particles, you will also receive purified pAAV-CAG-DIO-EYFP plasmid DNA.
CAG-driven, Cre-dependent expression of EYFP. These AAV were produced with the PHP.V1 serotype, which permits efficient transduction of brain vascular cells. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 7×10¹² 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, PHP.V1 cap gene pUCmini-iCAP-PHP.V1 (plasmid #127847)
- BufferPBS + 0.001% Pluronic F-68 + 200 mM NaCl
- SerotypePHP.V1
- PurificationIodixanol gradient ultracentrifugation
- Reporter GeneEYFP (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
- 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.
Citation Information: When using the PHP.V1 serotype in future publications, please acknowledge Viviana Gradinaru and cite Kumar et al., Nat Methods 17(5):541-550. Pubmed.ebiomall.com






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2、将校正测量开关扳在“校正”位置。
3、插接电源线,打开电源开关,并预热数分钟(待指针完全稳定下来为止)调节“调正”调节器使电表指示满度。
4、当使用(1)-(8)量程来测量电导率低于300μS•cm-1的液体时,选用“低周”,这时将高/低周开关扳向低周即可。当使用(9)-(10)量程来测量电导率在300μS•cm-1至105μS•cm-1范围里的液体时,则将扳向“高周”。
5、将量程选择开关扳到所需要的测量范围,如预先不知被测溶液电导率大小,应先把其扳到最大电导率测量档,然后逐渐下降,以防表针打弯。
6、电极的使用:使用时用电极夹夹紧电极的胶木帽,并把电极夹固定在电极杆上。
(1)当被测溶液的电导率低于0.3μS•cm-1,使用DJS-0.1型电极,这时应把“电极常数补偿调节器”调节在所配套电极常数的10倍位置上:例如,配套电极常数为0.090,则应把其调节到0.90位置上。
(2)当被测溶液的电导率低于10μS•cm-1,使用DJS-0.1型电极,这时应把“电极常数补偿调节器”调节在所配套电极常数相对应位置上:例如,配套电极常数为0.95,则应把其调节到0.95位置上,又若配套电极常数为1.1,则应调节在1.1位置上。
7、将电极插头插入电极插口内,旋紧插口上的紧固螺丝,再将电极綅入待测溶液中。
8、接着校正[当用(1)-(8)量程测量时,校正时扳到低周,当用(9)-(12)量程测量时,则校正扳到高周,扳到“校正”,调节校正调节器,使指示在满度。
9、当用(0-0.1)或(0-0.3)μS•cm-1这两档测量高纯水时,先把电极引线插入电极插孔,在电极未綅入溶液前,调节电容补偿调节器使电表指示为最小值(此最小值即电极铂片间的漏电阻,由于此漏电阻的存在,使得调电容补偿调节器时电表指针不能达到零点)。然后开始测量。
最近在装一根2L的层析柱(50×1000mm)出现了一个问题:
柱子装好后放置过夜总是出现了干柱现象。
我装柱的步骤是:
1、填料清洗(sephacrylS300),最后溶于ddH2O中。
2、空柱清洗加上装柱器,堵住下面的出水口,同时往柱子内灌入100mlddH2O中。
3、灌入填料匀浆。
4、打开下面出水口,用AKTAFPLC缓慢加压至目标反压。
5、堵住下出水口,旋入层析柱上接头并堵住出水口,4度放置。
这样子到了第二天总是出现了干柱的现象,我反复检查了出水口堵头并重装了几次,同样的问题依然出现。
我使用的层析柱是上海锦华层析设备厂的,不知前辈们可有遇到这样的问题,还望赐教!
具体的操作方法是怎样的,为什么我用的时候总是需要很长的时间才能稳定下来,我的测的纯化水的电导率和制水车间显示的有一定的差距,是测量的问题还是其他什么地方存在这种误差?
谢谢大家!

