Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Plasmid | 22222 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
| AAV1 | 22222-AAV1 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV9 | 22222-AAV9 | 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 backboneAAV-FLEX(Search Vector Database)
- Backbone manufacturerScott Sternson
- Backbone sizew/o insert(bp)5000
- Vector typeMammalian Expression, AAV, Cre/Lox ; adeno-associated virus
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature30°C
- Growth Strain(s)NEB Stable
- Growth instructionsUse recombinase-free E. coli (e.g., Invitrogen's Stbl3), grow at 30C
- Copy numberLow Copy
Gene/Insert
- Gene/Insert nameArch
- Alt nameArch-GFP
- Alt namearchaerhodopsin-3
- Alt nameH. sodomense archaerhodopsin-3
- SpeciesH. sodomense
- Insert Size (bp)1500
- Mutationno
- PromoterCAG
- Tag/ Fusion Protein
- GFP (C terminal on insert)
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteKpnI(not destroyed)
- 3′ cloning siteBsrGI(not destroyed)
- 5′ sequencing primerCCCATAACTTCGTATAATGTATGC (Common Sequencing Primers)
Resource Information
- Terms and Licenses
- UBMTA
- Ancillary Agreement for Plasmids Containing FP Materials
- genOway Notice of RIghts
- Industry Terms
- Not Available to Industry
- Articles Citing this Plasmid
- 15 References
Depositor Comments
PLEASE CONTACT ED BOYDEN ([email protected]) FOR DETAILS ON THIS REAGENT AND FURTHER REAGENTS IN THIS LINE.
In the map below, the Selected Features list should be labeled as follows (instead of the 3 loxP sites listed from 3629-3692 and 5299-5403): lox2272: 3629-3596 , loxp: 3725-3692 lox2272: 5266-5399 , loxp: 5370-5403
Information for AAV1 (Catalog # 22222-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from AAV-FLEX-Arch-GFP (#22222). In addition to the viral particles, you will also receive purified AAV-FLEX-Arch-GFP plasmid DNA.
CAG-driven cre-dependent Arch-GFP expression for optogenetic neural inhibition.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, AAV1 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV1
- PurificationIodixanol gradient ultracentrifugation
- Reporter GeneGFP (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.
Information for AAV9 (Catalog # 22222-AAV9)(Back to top)
Purpose
Ready-to-use AAV9 particles produced from AAV-FLEX-Arch-GFP (#22222). In addition to the viral particles, you will also receive purified AAV-FLEX-Arch-GFP plasmid DNA.
CAG-driven cre-dependent Arch-GFP expression for optogenetic neural inhibition.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, AAV9 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV9
- PurificationIodixanol gradient ultracentrifugation
- Reporter GeneGFP (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.
Data submitted about 22222-AAV9 by requesting scientist(s):
- Data 1: Mouse, Brain parenchyma
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QC实验室的电子天平配有打印机，早已不是什么业界的新闻了。瑞士梅特勒-托利多公司的和德国赛多利斯等国际知名电子天平生产商在N年前（不夸张地说，20年前）就给电子天平配置了打印机，并且电子天平也可以和计算机相连接，将称量数据保存在计算机中。
我国的很多大药企和经过FDA、欧盟认证的药企，基本上都为电子天平配置了打印机和（或者）计算机，将称量数据及时打印出来，和（或者0将称量数据存储在计算机中。
电子天平配置打印机的目的是为了将打印数据及时打印出来，防止称量数据被丢失和人为的改动，或者称量数据不原始（有很多人习惯先将称量数据记录在本上或者纸上，然后在记录在正式的记录上，这也是符合TCH和cGMP的要求的。
我们为电子天平配置上打印机就能做到称量数据真实了吗？我说不一定。当然主流是操作人员会把称量数据及时真实地打印出来。非主流还是可以对称量数据可以做手脚的。为什么这样说？因为如果实验室没有LIMS系统，那么你的电子天平是独立的，操作也是独立的，没有被监视和控制。因为称量操作也会发生失败（比如称量结果超出称量范围或者称量完必，在称量物质转移时发生洒落等现象）。操作人员势必会重新进行称量，那么上次称量的数据就很可能被删除或者不打印出来。
另外，称量的时间也会被人为地改变。一是改变电子天平上的时间设置，二是改变打印机和（或者）计算机上的时间来做假。
所以，我们在给电子天平配置了打印机和（或者）计算机后，我们还应该有合适的文件或者LIMS系统设置和规定和操作人员、主管等人的权限，使我们的称量操作符合法规的要求。避免和减少差错的发生，是GMP的核心内容之一。
以上内容，仅供同行们参考，如有不妥，欢迎指正。因个人水平有限，写的内容会不全，欢迎朋友们进行补充，坛友们会感谢您的。
(2) 向盘中加减砝码时要用镊子，不能用手接触砝码，不能把砝码弄湿、弄脏；（否则，砝码会沾上污物或被手上的汗水等锈蚀砝码，造成称量结果不精确）
(3) 潮湿的物体和化学物品不能直接放到天平盘中；（否则，会腐蚀天平托盘，减少天平使用寿命）
另外，还应注意：
(4) 搬动天平时，应轻拿轻放，不可让天平受到强烈振动；（否则，会损伤天平，造成天平的称量的灵敏度和精确度降低或减少天平的使用寿命）
(5) 用毕，应将游码归零，砝码归盒；横梁用固定圈固定，保证支架刀口不受损伤（此条适用于学生天平）；（否则，会损伤天平，造成天平的称量的灵敏度和精确度降低或减少天平的使用寿命。）
二个分度值的具体含义：
1.d是天平的最小刻度，称为分度值，也可叫作最小读数精度。按规定标准称其为天平实际标尺分度值，以符号“d”来代表。
2.e是天平检定标尺分度值，即检定时所用的分度值。检定时用的分度值和天平实际标尺分度值d不同，它不和d值一样是固定不变的，而是需要根据人为规定变化的值。可以规定e=5d，也可以规定e=10d，或者e=2d均可成立，只要不违反国家计量检定规程的规定就可以。
电子天平实际标尺分度值d是表示电子天平的最小精度是多少，检定标尺分度值e是电子天平进行检定时考核技术指标用的。准确的说，看电子天平的最佳精准度不能只看d值，还要看e值。特别是精确称量、科学实验及量值传递等部门在选择精密电子天平如0.01mg电子天平时最好要看检定标尺分度值e。
请问，临床前研究用的电子天平，需要符合GMP要求吗？需要3Q认证吗？相关数据用于申报临床试验的，谢谢
1251分析天平称重提到：电子分析天平有基于所用负载的内部校准系统。此校准适用于当前的环境温度。
<41>砝码与天平里面提到：2级砝码用于内置砝码的分析天平的校准工作，以及用于日常分析工作的实验室砝码。
但是看到一份培训资料，说美国fda不认可电子天平的自校系统的校验。
希望了解的战友执教，天平的自校和外置砝码校验是否都被fda接受。
谢谢
保存在巴黎的国际计量局总部国际千克原器，在2007年的一次检查中，发现有118年历史、用铂和铱混合铸造的圆柱形铸件——国际千克原器减轻了大约50毫克。2013年1月初，德国最新一期《计量学》杂志刊载研究报告称，作为标准质量单位的国际千克原器因表面遭污染而略有增重。
由此联想到FDA不承认内置砝码校准的问题，内置砝没有处在“千克原器”的真空、恒温、恒湿的环境，一年四季温度、环境湿度处在不断变化过程，降温和环境湿度增加时，总会有水分子凝结在内置砝码表面。升温和环境湿度下降时，总会有吸附在FDA内置砝码上的水分子解吸附，内置砝码自身的质量本身就是不断波动的过程，这样理解FDA不承认内置砝码的校准是否正确。请明白的战友指正。
我公司新购进的一台梅特勒XUS205U的十万分之一天平，他可以进行十万分之一与万分之一的转换，d值0.01mg/0.1mg跟随工程师做了四角偏差，重复性的测试，关于最小称样量工程师拒绝给做。想请教各位有没有知道怎么做的。目前了解的就是用1g的砝码做标准偏差然后反推，但是还不是很清楚怎么做
