Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Plasmid | 50474 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
| AAV2 | 50474-AAV2 | Virus (100 µL at titer ≥ 3×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV5 | 50474-AAV5 | Virus (100 µL at titer ≥ 3×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV8 | 50474-AAV8 | Virus (100 µL at titer ≥ 2×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV Retrograde | 50474-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 backbonepAAV(Search Vector Database)
- Backbone sizew/o insert(bp)4818
- Total vector size (bp)7070
- Vector typeAAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)NEB Stable
- Copy numberLow Copy
Gene/Insert
- Gene/Insert namehM3D(Gq)-mCherry
- SpeciesH. sapiens (human)
- Insert Size (bp)2502
- MutationSee supplemental documents for DREADD mutations
- Entrez GeneCHRM3(a.k.a. EGBRS, HM3, PBS)
- Promoterhuman Synapsin 1
- Tag/ Fusion Protein
- mCherry (C terminal on insert)
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteSal I(not destroyed)
- 3′ cloning siteEcoR I(not destroyed)
- 5′ sequencing primerTCGTGTCGTGCCTGAGAGCG
- 3′ sequencing primerGCATTAAAGCAGCGTATCCACATAGC (Common Sequencing Primers)
Resource Information
- Supplemental Documents
- DREADD mutations
- Terms and Licenses
- UBMTA
- Takara Bio Limited Use Label License (formerly Clontech)
- Industry Terms
- Not Available to Industry
- Articles Citing this Plasmid
- 5 References
Depositor Comments
Please Note- This plasmid does NOT contain an N-terminal HA tag.
For more information on DREADDs, see: http://pdspit3.mml.unc.edu/projects/dreadd/wiki/WikiStart
Information for AAV2 (Catalog # 50474-AAV2)(Back to top)
Purpose
Ready-to-use AAV2 particles produced from pAAV-hSyn-hM3D(Gq)-mCherry (#50474). In addition to the viral particles, you will also receive purified pAAV-hSyn-hM3D(Gq)-mCherry plasmid DNA.
hSyn-driven hM3D(Gq) receptor with an mCherry reporter for CNO-induced neuronal activation. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 3×10¹² 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
- PurificationCsCl gradient ultracentrifugation
- Reporter GenemCherry
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
Data submitted about 50474-AAV2 by requesting scientist(s):
- Data 1: Rat, Brain parenchyma
Information for AAV5 (Catalog # 50474-AAV5)(Back to top)
Purpose
Ready-to-use AAV5 particles produced from pAAV-hSyn-hM3D(Gq)-mCherry (#50474). In addition to the viral particles, you will also receive purified pAAV-hSyn-hM3D(Gq)-mCherry plasmid DNA.
hSyn-driven hM3D(Gq) receptor with an mCherry reporter for CNO-induced neuronal activation. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 3×10¹² 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 GenemCherry
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.
Information for AAV8 (Catalog # 50474-AAV8)(Back to top)
Purpose
Ready-to-use AAV8 particles produced from pAAV-hSyn-hM3D(Gq)-mCherry (#50474). In addition to the viral particles, you will also receive purified pAAV-hSyn-hM3D(Gq)-mCherry plasmid DNA.
hSyn-driven hM3D(Gq) receptor with an mCherry reporter for CNO-induced neuronal activation. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 2×10¹² 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, AAV8 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV8
- PurificationIodixanol gradient ultracentrifugation
- Reporter GenemCherry
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.
Information for AAV Retrograde (Catalog # 50474-AAVrg)(Back to top)
Purpose
Ready-to-use AAV Retrograde particles produced from pAAV-hSyn-hM3D(Gq)-mCherry (#50474). In addition to the viral particles, you will also receive purified pAAV-hSyn-hM3D(Gq)-mCherry plasmid DNA.
hSyn-driven hM3D(Gq) receptor with an mCherry reporter for CNO-induced neuronal activation. 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 GenemCherry
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.Data submitted about 50474-AAVrg by requesting scientist(s):
- Data 1: Rat, Brain parenchyma
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② 应取pH8.0,这样可使核苷酸带较多负电荷,利于吸附于阴离子交换树脂柱。虽然pH 11.4时核苷酸带有更多的负电荷,但pH过高对分离不利。
③ 当不考虑树脂的非极性吸附时,根据核苷酸负电荷的多少来决定洗脱速度,则洗脱顺序为CMP>AMP> GMP > UMP,但实际上核苷酸和聚苯乙烯阴离子交换树脂之间存在着非极性吸附,嘌呤碱基的非极性吸附是嘧啶碱基的3倍。静电吸附与非极性吸附共同作用的结果使洗脱顺序为:CMP> AMP > UMP >GMP。
1. Buffer中离子浓度过大,甘氨酸或者Tris碱有可能疏忽多加了
2.没有加相应浓度的SDS
3.电泳周围温度高,也是一个原因
我使用的是1%琼脂糖凝胶,电泳缓冲液为0.5×TBE。
请问各位大虾,在不影响实验结果的前提下,这样的含EB的电泳缓冲液可以反复使用多少次?
谢谢^_^
500ml
2。阴极缓冲液 ( 10 × ) :将 60.55g Tris ,89.58g Tricine 及 5g SDS 溶于400ml 蒸馏水中,加水至终体积为500ml
使用时稀释至1×的缓冲液,电泳槽内槽加入阴极电泳缓冲液,外槽加入阳极电泳缓冲液。形成Trcine-SDS-PAGE电泳系统。
如果你是制胶或者做电泳缓冲液的话,不用灭菌。
Tris-Glycine/SDS;
MOPS/SDS;
Bis-Glycine/SDS等不同的缓冲液、预混液等。
不知道这几种缓冲液有什么异同呢?
比如如果是Invitrogen的预制胶-预混液电泳系统,每种预制胶有适配缓冲液,但是如果用其他的缓冲液似乎也可以得到不错的结果,那么是否说明电泳缓冲液只要满足了缓冲ph范围,其他的不是特别重要呢?
此外,附加提问是,PVDF膜在转膜的时候,转膜缓冲液中不需要加入甲醇,那么是加入甲醇转膜效果好还是不加好呢?
谢谢!

