
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Plasmid | 104493 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
AAV1 | 104493-AAV1 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
AAV Retrograde | 104493-AAVrg | 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 backboneAAV-Syn-FLEX(Search Vector Database)
- Backbone manufacturerScott Sternson
- Backbone sizew/o insert(bp)4894
- Total vector size (bp)6247
- Vector typeMammalian Expression, AAV, Cre/Lox
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature30°C
- Growth Strain(s)NEB Stable
- Copy numberLow Copy
Gene/Insert
- Gene/Insert namejGCaMP7b
- Alt nameGCaMP3-T302P R303P A317L M374Y D380Y T381R S383T R392G
- Alt nameGCaMP3 variant 1561
- Alt nameJanelia GCaMP7
- SpeciesR. norvegicus (rat), G. gallus (chicken); A. victoria (jellyfish)
- Insert Size (bp)1353
- PromoterSynapsin
- Tag/ Fusion Protein
- T7 epitope, Xpress tag, 6xHis
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteBsmBI(destroyed during cloning)
- 3′ cloning siteBsmBI(destroyed during cloning)
- 5′ sequencing primerACCACGCGAGGCGCGAGATAG (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
Information for AAV1 (Catalog # 104493-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from pGP-AAV-syn-FLEX-jGCaMP7b-WPRE (#104493). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7b-WPRE plasmid DNA.
Synapsin-driven, Cre-dependent jGCaMP7b expression. jGCaMP7b exhibits the brightest resting fluorescence and can be used for imaging of small neuronal processes (dendrites and axons). 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
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 AAV Retrograde (Catalog # 104493-AAVrg)(Back to top)
Purpose
Ready-to-use AAV Retrograde particles produced from pGP-AAV-syn-FLEX-jGCaMP7b-WPRE (#104493). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7b-WPRE plasmid DNA.
Synapsin-driven, Cre-dependent jGCaMP7b expression. jGCaMP7b expression exhibits the brightest resting fluorescence and can be used for imaging of small neuronal processes (dendrites and axons). These AAV were produced with a retrograde serotype, which permits retrograde access to projection neurons. 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, AAV retrograde cap gene from rAAV2-retro helper (plasmid #81070)
- BufferPBS + 0.001% Pluronic F-68 + 200 mM NaCl
- SerotypeAAV retrograde (AAVrg)
- PurificationIodixanol gradient ultracentrifugation
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
Retrograde functionality is dependent on high viral titers. Addgene recommends not diluting your AAV preps prior to use.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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成像原理:
光学显微镜
光学显微镜主要由目镜、物镜、载物台和反光镜组成。目镜和物镜都是凸透镜,焦距不同。物镜的凸透镜焦距小于目镜的凸透镜的焦距。物镜相当于投影仪的镜头,物体通过物镜成倒立、放大的实像。目镜相当于普通的放大镜,该实像又通过目镜成正立、放大的虚像。经显微镜到人眼的物体都成倒立放大的虚像。反光镜用来反射,照亮被观察的物体。反光镜一般有两个反射面:一个是平面镜,在光线较强时使用;一个是凹面镜,在光线较弱时使用,可会聚光线。
电子显微镜
电子显微镜是根据电子光学原理,用电子束和电子透镜代替光束和光学透镜,使物质的细微结构在非常高的放大倍数下成像的仪器。
电子显微镜的分辨能力以它所能分辨的相邻两点的最小间距来表示。20世纪70年代,透射式电子显微镜的分辨率约为0.3纳米(人眼的分辨本领约为0.1毫米)。现在电子显微镜最大放大倍率超过300万倍,而光学显微镜的最大放大倍率约为2000倍,所以通过电子显微镜就能直接观察到某些重金属的原子和晶体中排列整齐的原子点阵。
详见百度百科:http://baike.baidu.com/view/2921.htm
1,先把视频显微镜的几个大的零件准备好,先把万向支架组装起来,在这里要注意镜头和相机的支架应该在下面,支持显示器的支架要放到上面,这样便于观察。
2,把镜头安装在万向支架上面,安装到有一个圈的里面,正好固定镜头,镜头是比较脆弱的,所以安装的时候千万要小心,然后再把相机安装到镜头的上面,哪里有接口直径插上就可以了,这一步是很重要的,千万小小心。
3,支架和镜头安装好以后然后把显示器固定好,固定好以后,再把一些链接线连上这样就可以了。
总放大倍数 = 物镜放大倍数 * 数字放大倍数
物镜放大倍数 = 大物镜放大倍数 * 镜头放大倍数
数字放大倍数 = 监视器尺寸 * 25.4/CCD靶面对角线尺寸大小
CCD靶面对角线尺寸大小:1/3 " 为6mm1/2 " 为8mm 2/3 " 为11mm
例:0.7X - 4.5X的标配主机配1/3 "CCD摄像机配14 " 监视器
数字放大倍数:14 * 25.4 / 6 = 59.3X
总放大倍数:(0.7X - 4.5X) * 59.3=41.5X - 266.9X
那么照此配置,总的放大倍率就在41.5X到266.9X之间连续可调。
LG液晶热点侦测:利用液晶感测到IC漏电处分子排列重组,在显微镜下呈现出不同于其它区域的斑状影像,找寻在实际分析中困扰设计人员的漏电区域(超过10mA之故障点)。
定点/非定点芯片研磨:移除植于液晶驱动芯片 Pad上的金凸块, 保持Pad完好无损,以利后续分析或rebonding。
X-Ray 无损侦测:检测IC封装中的各种缺陷如层剥离、爆裂、空洞以及打线的完整性,PCB制程中可能存在的缺陷如对齐不良或桥接,开路、短路或不正常连接的缺陷,封装中的锡球完整性。
列文虎克又动手做了一个金属支架和一个小圆筒,把两块镜片分别装在圆筒两头,还安上旋钮,来调节两块镜片间的距离。这样,世界上第一台显微镜就诞生了。

