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Addgene/pGP-AAV-syn-FLEX-jGCaMP7s-WPRE/1unit/104491-PHPeB

价格
¥7600.00
货号:104491-PHPeB
浏览量:111
品牌:Addgene
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商品描述

Ordering

ItemCatalog #DescriptionQuantityPrice (USD)
Plasmid104491Standard format: Plasmid sent in bacteria as agar stab1$75
Add to Cart
AAV1104491-AAV1100 µL at titer ≥ 1×10¹³ vg/mLand Plasmid.More Information
$380
Add to Cart
AAV9104491-AAV9100 µL at titer ≥ 1×10¹³ vg/mLand Plasmid.More Information
$380
Add to Cart
AAV Retrograde104491-AAVrgVirus (100 µL at titer ≥ 7×10¹² vg/mL)and Plasmid.More Information
$380
Add to Cart
AAV PHP.eB104491-PHPeB100 µL at titer ≥ 1×10¹³ vg/mLand Plasmid.More Information
$380
Add to Cart

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    AAV-Syn-FLEX
    (Search Vector Database)
  • Backbone manufacturer
    Scott Sternson
  • Backbone sizew/o insert(bp)4894
  • Total vector size (bp)6247
  • Vector type
    Mammalian Expression, AAV, Cre/Lox

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    30°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    jGCaMP7s
  • Alt name
    GCaMP3-A52V K78H T302L R303P A317L D380Y T381R S383T R392G
  • Alt name
    GCaMP3 variant 1473
  • Species
    R. 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
  • Article Citing this Plasmid
    • 1 Reference

Information for AAV1 (Catalog # 104491-AAV1)(Back to top)

Purpose

Ready-to-use AAV1 particles produced from pGP-AAV-syn-FLEX-jGCaMP7s-WPRE (#104491). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7s-WPRE plasmid DNA.

Synapsin-driven, Cre-dependent, GCaMP7s calcium sensor.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

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 # 104491-AAV9)(Back to top)

Purpose

Ready-to-use AAV9 particles produced from pGP-AAV-syn-FLEX-jGCaMP7s-WPRE (#104491). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7s-WPRE plasmid DNA.

Synapsin-driven, Cre-dependent, GCaMP7s calcium sensor.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

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

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 # 104491-AAVrg)(Back to top)

Purpose

Ready-to-use AAV Retrograde particles produced from pGP-AAV-syn-FLEX-jGCaMP7s-WPRE (#104491). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7s-WPRE plasmid DNA.

Synapsin-driven, Cre-dependent, GCaMP7s calcium sensor.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

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.

Information for AAV PHP.eB (Catalog # 104491-PHPeB)(Back to top)

Purpose

Ready-to-use AAV PHP.eB particles produced from pGP-AAV-syn-FLEX-jGCaMP7s-WPRE (#104491). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7s-WPRE plasmid DNA.

Synapsin-driven, Cre-dependent, GCaMP7s calcium sensor.These AAV were produced with the PHPeB serotype, which permits efficient transduction of the central nervous system. 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, PHP.eB cap gene pUCmini-iCAP-PHP.eB (plasmid #103005)
  • BufferPBS + 0.001% Pluronic F-68
  • SerotypePHPeB (plasmid #103005)
  • 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

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.eB serotype in future publications, please acknowledge Viviana Gradinaru and cite Chan et al., Nat Neurosci, 20(8):1172-1179. Pubmed.
Addgene所述Ç ircular p ermutation基于˚F luorescence ř esonance Ë NERGY 吨转让(BOT)(cpFRET)生物传感器工具包是用于产生和/或遗传编码,单分子FRET传感器,用于比例测量的优化的载体文库。该工具包旨在简化和加速生物传感器的生产,面向对信号转导过程的时空分析感兴趣的广大社区。cpFRET试剂盒的标志是影响FRET效率的参数的变化:荧光团之间的距离由不同长度的接头修饰,而荧光团的偶极子方向则通过在mTFP1供体和金星受体荧光团中引入的圆形排列来操纵。另外,可以使用不同的感测模块拓扑来进一步增强几何多样性。这也使得能够容纳需要天然N或C末端用于脂质修饰的信号传导分子。这个基于pTriEx的库由两个设计组成,每个设计包含25个生物传感器。pTriEx载体系统可在大肠杆菌,昆虫和脊椎动物细胞中表达生物传感器。在一种设计中,将野生型(wt)mTFP1荧光团及其四个圆形排列与野生型金星荧光团及其四个圆形排列组合在一起。设计1生物传感器根据外部荧光团构建:mTFP1(wt和cp变体)后接脯氨酸导向的WW磷酸结合结构域,72个氨基酸(aa)甘氨酸接头,10个氨基酸的ERK底物肽Cdc25C,一个四氨基酸ERK对接位点,金星(wt和cp变体)和一个核出口信号(图1A)。设计2传感器是根据荧光团内部结构构建的:WW域后是mTFP1(wt和cp变体),20 aa接头,金星(wt和cp变体),Cdc25C底物肽,ERK对接位点和核出口信号(图1B)。通过简单的域交换生成设计3和4可以生成两个其他设计,它们显示了混合结构,其中荧光团与传感模块组件交替出现(图1C)。为了生产新的生物传感器或优化已经存在的生物传感器,可以通过选择的新传感模块交换WW域和ERK底物/ ERK对接肽。可以去除核出口信号以构造核报告仪。此外,荧光团也可以被取代,以利用新的供体/受体组合。由于独特的限制位点位于生物传感器模块中所有组件的两侧,因此简化了克隆过程。用感兴趣的传感模块生产了一个新的文库后,可以使用一种表现出稳定的生物传感器ON和OFF状态的检测方法筛选整个文库,以鉴定具有最高动态范围和最理想的光谱特性的传感器。