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Addgene/pAAV-CAG-tdTomato (codon diversified)/1unit/59462-PHPeB
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4000-520-616
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Plasmid | 59462 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
| AAV1 | 59462-AAV1 | Virus (100 µL at titer ≥ 5×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV2 | 59462-AAV2 | Virus (100 µL at titer ≥ 7×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV5 | 59462-AAV5 | Virus (100 µL at titer ≥ 5×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV Retrograde | 59462-AAVrg | Virus (100µL at titer ≥ 7×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV PHP.S | 59462-PHP.S | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV PHP.eB | 59462-PHPeB | 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 with CAG promter(Search Vector Database)
- Backbone manufacturerScott Sternson
- Backbone sizew/o insert(bp)4719
- Total vector size (bp)6150
- Vector typeMammalian Expression, AAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)NEB Stable
- Growth instructionsStbl3 at 30C (use carbenicillin if using Stbl3) OR DH5a at 37C
- Copy numberLow Copy
Gene/Insert
- Gene/Insert nametdTomato
- SpeciesSynthetic
- Insert Size (bp)1431
- Mutationcodon diversified
- PromoterCAG
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteBamHI(not destroyed)
- 3′ cloning siteEcoRI(not destroyed)
- 5′ sequencing primergctctagagcctctgctaacc
- 3′ sequencing primergcagcgtatccacatagcg (Common Sequencing Primers)
Resource Information
- Terms and Licenses
- UBMTA
- Takara Bio Limited Use Label License (formerly Clontech)
- Industry Terms
- Not Available to Industry
- Articles Citing this Plasmid
- 7 References
Depositor Comments
Plasmid is completely sequenced by depositing lab except for parts of both ITRs and a part of the CAG promoter. Multiple digestions were done to verify the vector structure. The construct and the virus were both tested in vitro.
Information for AAV1 (Catalog # 59462-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from pAAV-CAG-tdTomato (codon diversified) (#59462). In addition to the viral particles, you will also receive purified pAAV-CAG-tdTomato (codon diversified) plasmid DNA.
CAG-driven tdTomato expression control. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 5×10¹² 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, AAV1 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV1
- PurificationIodixanol gradient ultracentrifugation
- Reporter GenetdTomato (codon diversified)
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.
Information for AAV2 (Catalog # 59462-AAV2)(Back to top)
Purpose
Ready-to-use AAV2 particles produced from pAAV-CAG-tdTomato (codon diversified) (#59462). In addition to the viral particles, you will also receive purified pAAV-CAG-tdTomato (codon diversified) plasmid DNA.
CAG-driven tdTomato expression control. 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, AAV2 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV2
- PurificationIodixanol gradient ultracentrifugation
- Reporter GenetdTomato (codon diversified)
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.
Information for AAV5 (Catalog # 59462-AAV5)(Back to top)
Purpose
Ready-to-use AAV5 particles produced from pAAV-CAG-tdTomato (codon diversified) (#59462). In addition to the viral particles, you will also receive purified pAAV-CAG-tdTomato (codon diversified) plasmid DNA.
CAG-driven tdTomato expression control. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 5×10¹² 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, AAV5 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV5
- PurificationIodixanol gradient ultracentrifugation
- Reporter GenetdTomato (codon diversified)
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.
Information for AAV Retrograde (Catalog # 59462-AAVrg)(Back to top)
Purpose
Ready-to-use AAV Retrograde particles produced from pAAV-CAG-tdTomato (codon diversified) (#59462). In addition to the viral particles, you will also receive purified pAAV-CAG-tdTomato (codon diversified) plasmid DNA.
tdTomato expression control. 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
- Reporter GenetdTomato (codon diversified)
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.Data submitted about 59462-AAVrg by requesting scientist(s):
- Data 1: Mouse, Brain parenchyma
- Data 2: Mouse, Cerebrospinal fluid (CSF): Intracerebroventricular (ICV)
- Data 3: Mouse, Cerebrospinal fluid (CSF): Intracerebroventricular (ICV)
- Data 4: Mouse, Brain parenchyma
Information for AAV PHP.S (Catalog # 59462-PHP.S)(Back to top)
Purpose
Ready-to-use AAV PHP.S particles produced from pAAV-CAG-tdTomato (codon diversified) (#59462). In addition to the viral particles, you will also receive purified pAAV-CAG-tdTomato (codon diversified) plasmid DNA.
CAG-driven tdTomato expression control. These AAV were produced with the PHP.S serotype, which permits efficient transduction of the peripheral 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.S cap gene pUCmini-iCAP-PHP.S (plasmid #103006)
- BufferPBS + 0.001% Pluronic F-68
- SerotypePHP.S (plasmid #103006)
- PurificationIodixanol gradient ultracentrifugation
- Reporter GenetdTomato (codon diversified)
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
Citation Information: When using the PHP.S serotype in future publications, please acknowledge Viviana Gradinaru and cite Chan et al., Nat Neurosci, 20(8):1172-1179. Pubmed.Data submitted about 59462-PHP.S by requesting scientist(s):
- Data 1: Mouse, Intravenous (retro-orbital or tail vein)
Information for AAV PHP.eB (Catalog # 59462-PHPeB)(Back to top)
Purpose
Ready-to-use AAV PHP.eB particles produced from pAAV-CAG-tdTomato (codon diversified) (#59462). In addition to the viral particles, you will also receive purified pAAV-CAG-tdTomato (codon diversified) plasmid DNA.
CAG-driven tdTomato expression control. 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
- Reporter GenetdTomato (codon diversified)
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
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.Data submitted about 59462-PHPeB by requesting scientist(s):
- Data 1: Mouse, Intravenous (retro-orbital or tail vein)
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【求助/交流】dna测浓度怎样选择合适的稀释倍数 123
kvza5052014-12-12
大家好，我们实验室有一种酶活为30KU每ml的Dnase1，由于其酶活太高，所以想进行稀释后使用，但是由于没有说明书，不知道如何稀释，请问应该如何稀释呢？稀释液的配方如何配制？有什么要注意的吗？谢谢
【求助】基因组dna酶切 核酸基因技术讨论版论坛123
nestea1102008-11-25
最近准备做southern，但是提取的苜蓿基因组dna酶切总是不尽如人意
同一管dna有的酶可以切开，比如dra1，ecoR1,Hind111
有的酶又完全不能切动，如BamH1，sac1，xba1，xho1，pst1
请问高手这是什么原因？?
同一管dna有的酶可以切开，比如dra1，ecoR1,Hind111
有的酶又完全不能切动，如BamH1，sac1，xba1，xho1，pst1
请问高手这是什么原因？?
实验九DNA的酶切123
园香丁dingxiang2016-09-01
求问酶切体系为10ul,内切酶1ul,37℃水浴45min，不知道DNA产物是否完全被切开了？
所用内切酶信息如下
艾弗里实验中为什么最后要增加将S型细菌的DNA和DNA酶一起加入R型细菌这...123
2017-09-29
高中生物必修二中，艾弗里实验最后一组实验为什么要加DNA和DNA酶到R型细菌里？
dna聚合酶ⅲ全酶结构全酶结构包括.ppt123
湛之吻2017-09-29
DNA即脱氧核糖核酸（deoxyribonucleicacid）,其基本组成单位是核苷酸，而核苷酸又是有核酸，戊糖和磷酸组成（DNA中的戊糖为脱氧核糖）。DNA水解酶为一类可以将组成DNA分子的脱氧核糖核苷酸之间的连接（3’，5’-磷酸二酯键）打开的酶。具体包括：DNA聚合酶α/引发酶（引发及后随链的部分合成），DNA聚合酶δ（DNA复制主要酶），增殖细胞核抗原（滑动夹子，与合成连接性有关），拓扑异构酶（母链DNA拓扑异构化），解（螺）旋酶（能解开DNA双螺旋），单链DNA结合蛋白及复制蛋白（单链DNA结合作用），复制因子C（参与滑动夹子的装配），DNA连接酶（连接冈崎片段及参与修复），核酸酶（去除RNA引物），侧翼核酸内切酶（去除RNA引物）。总体作用是在DNA复制过程中发挥的，在DNA复制过程中起到关键性的作用。
【求助】DNA酶切、连接的困惑_问答详情_限制性内切酶_核酸酶类_...123
2547747552010-03-22
各位大侠小弟有一事很是郁闷，请各位赐教！
我做的双酶切反应，酶分别是宝生物的BamHI和XhoI，双酶切体系是：载体及目的片段分别是30ul，通用buffer4ul，XhoI、BamHI各2ul，无核酶水2ul，总体积40ul。载体和目的片段都是经37度酶切6h。载体上面这两个酶切位点的距离是6个碱基，目的片段是由pcr反应获得的，从puc19中p出来的，两端带有这两种酶的酶切位点，酶切完成后，做连接反应，体系如下：目的片段（全长1441bp）4ul，无核酶水3ul，10*T4DNA连接酶缓冲液1ul，载体1ul，T4DNA连接酶1ul，总体积10ul。16度过夜。之后摇菌送沉菌测序结果回来，送了5管一个都不对。重复实验两次还是不对！
之后改为先用BanHI单切载体，体系如下：载体17ul，酶1ul，buffer2ul，总体积20ul，30度切6小时后，Promega纯化试剂盒直接纯化，完了之后再用XhoI，37度酶切6小时，体系如上，再次用promega纯化试剂盒纯化，目的片段用双酶切体系酶切，之后做连接反应，之后铺板，挑取菌落共10个，摇菌13小时，取菌液做pcr鉴定，结果10个菌落全是引物2聚体，跑出来的条带都是100bp左右。
各位老师这是怎么回事呢？为什么连接不上呢？小弟先谢过各位大哥了！
我做的双酶切反应，酶分别是宝生物的BamHI和XhoI，双酶切体系是：载体及目的片段分别是30ul，通用buffer4ul，XhoI、BamHI各2ul，无核酶水2ul，总体积40ul。载体和目的片段都是经37度酶切6h。载体上面这两个酶切位点的距离是6个碱基，目的片段是由pcr反应获得的，从puc19中p出来的，两端带有这两种酶的酶切位点，酶切完成后，做连接反应，体系如下：目的片段（全长1441bp）4ul，无核酶水3ul，10*T4DNA连接酶缓冲液1ul，载体1ul，T4DNA连接酶1ul，总体积10ul。16度过夜。之后摇菌送沉菌测序结果回来，送了5管一个都不对。重复实验两次还是不对！
之后改为先用BanHI单切载体，体系如下：载体17ul，酶1ul，buffer2ul，总体积20ul，30度切6小时后，Promega纯化试剂盒直接纯化，完了之后再用XhoI，37度酶切6小时，体系如上，再次用promega纯化试剂盒纯化，目的片段用双酶切体系酶切，之后做连接反应，之后铺板，挑取菌落共10个，摇菌13小时，取菌液做pcr鉴定，结果10个菌落全是引物2聚体，跑出来的条带都是100bp左右。
各位老师这是怎么回事呢？为什么连接不上呢？小弟先谢过各位大哥了！
【求助】提取的RNA,DNA酶消化1小时为何还能扩出目的片段? PCR技...123
leukocyte2007-06-08
【请问】dna酶1和rna酶A可以用milliQ水[配制溶液吗? 形态学与...123
国宝2005-07-18
请问dna酶1和rna酶A可以用milliQ水[配制溶液吗?
配后是否应该分装,因为不能反复冻融?
多谢!
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【】可以这样输入，智能ABC，输入“v1”，后翻3页，就可以看见了，或用紫光拼音中文输入法，输入方括号就是粗体的中括号，实在不行请复制本公告中【】
配后是否应该分装,因为不能反复冻融?
多谢!
1、发帖请归入相应子版（选择SubBoard）！
2、请在帖子前面注明帖子性质，如【求助】、【原创】、【分享】、【讨论】、【申请上传】、【下载】、【转帖】、【转载】、【信息】、【求购】、【公告】、【建议】等，以方便战友应助及版面管理！
3、不规范的发帖及应助帖将不加分！
4、对于得分在5分以下或贴子数少于20的战友的不规范发贴，第一次我们将帮助修改同时给予提示；对于5分以上或贴子数大于20的战友的不规范发贴将直接锁贴，等待修改（可PM版主），保留24h仍然不修改，将删除所发的贴子而且不另外通知！
【】可以这样输入，智能ABC，输入“v1”，后翻3页，就可以看见了，或用紫光拼音中文输入法，输入方括号就是粗体的中括号，实在不行请复制本公告中【】
【求助】有人复苏细胞,去除DMSO后,用DNA酶孵育的吗??或者知道...123
elevenli2015-06-12
看文献,看到其复苏脐带血单个核细胞，去除DMSO后，用含有DNA酶的培养液孵育了一会。请问人在复苏细胞的时候也有类似的做法，或者知道这样做的具体原因吗？
感谢赐教~！
感谢赐教~！
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