
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Plasmid | 105530 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
AAV1 | 105530-AAV1 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
AAV2 | 105530-AAV2 | Virus (100 µL at titer ≥ 7×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
AAV5 | 105530-AAV5 | Virus (100 µL at titer ≥ 7×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
AAV8 | 105530-AAV8 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
AAV9 | 105530-AAV9 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
AAV rh10 (similar to AAV10) | 105530-rh10 | 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 backbonepAAV(Search Vector Database)
- Vector typeMammalian Expression, AAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)NEB Stable
- Copy numberUnknown
Gene/Insert
- Gene/Insert nameEGFP
- PromoterCMV
Cloning Information
- Cloning methodUnknown
- 5′ sequencing primerCMV-F (Common Sequencing Primers)
Resource Information
- Supplemental Documents
- pAAV.CMV.PI.EGFP.WPRE.bGH (p0101).gb
- Terms and Licenses
- UBMTA
- Ancillary Agreement for Plasmids Containing FP Materials
- Industry Terms
- Not Available to Industry
- Article Citing this Plasmid
- 1 Reference
Depositor Comments
Penn Vector Core number p0101
Information for AAV1 (Catalog # 105530-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from pAAV.CMV.PI.EGFP.WPRE.bGH (#105530). In addition to the viral particles, you will also receive purified pAAV.CMV.PI.EGFP.WPRE.bGH plasmid DNA.
CMV-driven EGFP control. 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 GeneEGFP
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 AAV2 (Catalog # 105530-AAV2)(Back to top)
Purpose
Ready-to-use AAV2 particles produced from pAAV.CMV.PI.EGFP.WPRE.bGH (#105530). In addition to the viral particles, you will also receive purified pAAV.CMV.PI.EGFP.WPRE.bGH plasmid DNA.
CMV-driven EGFP 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 GeneEGFP
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 AAV5 (Catalog # 105530-AAV5)(Back to top)
Purpose
Ready-to-use AAV5 particles produced from pAAV.CMV.PI.EGFP.WPRE.bGH (#105530). In addition to the viral particles, you will also receive purified pAAV.CMV.PI.EGFP.WPRE.bGH plasmid DNA.
CMV-driven EGFP 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, AAV5 cap gene
- BufferPBS + 0.001% Pluronic F-68 + 200 mM NaCl
- SerotypeAAV5
- PurificationIodixanol gradient ultracentrifugation
- Reporter GeneEGFP
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 # 105530-AAV8)(Back to top)
Purpose
Ready-to-use AAV8 particles produced from pAAV.CMV.PI.EGFP.WPRE.bGH (#105530). In addition to the viral particles, you will also receive purified pAAV.CMV.PI.EGFP.WPRE.bGH plasmid DNA.
CMV-driven EGFP control. 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, AAV8 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV8
- PurificationIodixanol gradient ultracentrifugation
- Reporter GeneEGFP
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 AAV9 (Catalog # 105530-AAV9)(Back to top)
Purpose
Ready-to-use AAV9 particles produced from pAAV.CMV.PI.EGFP.WPRE.bGH (#105530). In addition to the viral particles, you will also receive purified pAAV.CMV.PI.EGFP.WPRE.bGH plasmid DNA.
CMV-driven EGFP control. 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 GeneEGFP
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 rh10 (similar to AAV10) (Catalog # 105530-rh10)(Back to top)
Purpose
Ready-to-use AAV rh10 (similar to AAV10) particles produced from pAAV.CMV.PI.EGFP.WPRE.bGH (#105530). In addition to the viral particles, you will also receive purified pAAV.CMV.PI.EGFP.WPRE.bGH plasmid DNA.
CMV-driven EGFP control. 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, AAVrh10 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAVrh10
- PurificationIodixanol gradient ultracentrifugation
- Reporter GeneEGFP
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.
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1机械法:主要通过机械切力的作用使组织细胞破碎的方法,常用的器械有组织捣碎机、匀浆器、研钵和研磨、压榨器等。
1.1组织捣碎机
将材料配成稀糊状液,放置于筒内约1/3体积,盖紧筒盖,将调速器先拨至最慢处,开动开关后,逐步加速至所需速度。一般用于动物组织、植物肉质种子、柔嫩的叶芽等,转速可高达10000rpm/M以上。由于旋转刀片的机械切力很大,制备一些较大分子如核酸则很少使用。
1.2匀浆器
先将剪碎的组织置于管中,再套入研杆来回研磨,上下移动,即可将细胞研碎。匀浆器的研钵磨球和玻璃管内壁之间间隙保持在十分之几毫米距离。制作匀浆器的材料,除玻璃外,还可以用硬质塑料、不锈钢、人造荧光树脂等。此法细胞破碎程度比高速组织捣碎机为高,适用于量少和动物脏器组织。
存在的问题;较易造成堵塞的团状或丝状真菌,较小的革兰氏阳性首以及有些亚细胞器,质地坚硬,易损伤匀浆阀,也不适合用该法处理。
1.3. 研钵
多用于细菌或其他坚硬植物材料,研磨时常加入少量石英砂,玻璃粉或其他研磨剂,以提高研磨效果。
1.44. 细菌磨
是一种改良了的研磨器,比研钵具有更大的研磨面积,而且低部有出口。操作时先把细菌和研磨粉调成糊状,每次加入一小勺,研磨20-30秒即可将细菌细胞完全磨碎。
2 物理法
主要通过各种物理因素使组织细胞破碎的方法。在生化制备中常用的方法有:
2.1. 反复冻溶法
原理:因突然冷冻,细胞内冰晶的形成及胞内外溶剂浓度的突然改变而破坏细胞。
方法:将待破碎的细胞在-20度以下冰冻,室温融解,反复几次,由于细胞内冰粒形成和剩余细胞液的盐浓度增高引起溶胀,使细胞结构破碎。
特点:此法适用于组织细胞,多用于动物性材料,对微生物细胞作用较差。
2.2. 急热骤冷法
将材料投入沸水中,维持85-90分钟,至水浴中急速冷却,此法可用于细菌及病毒材料。
2.3. 超声波处理
用一定功率的超声波处理细胞悬液,使细胞急剧震荡破裂,此法多适用于微生物材料,用大肠杆菌制备各种酶,常选用50-100毫克菌体/毫升浓度,频高于15~20KHz的超声波在高强度声能输入下可以进行细胞破碎。其破碎机理:可能与空化现象引起的冲击波和剪切力有关。超声破碎的效率与声频、声能、处理时间、细胞浓度及首种类型等因素有关。
行细胞破碎。其破碎机理:可能与空化现象引起的冲击波和剪切力有关。超声破碎的效率与声频、声能、处理时间、细胞浓度及首种类型等因素有关。
特点:操作简单,重复性较好,节省时间;多用于微生物和组织细胞的破碎。
存在问题:超声波破碎在实验室规模应用较普遍,处理少量样品时操作简便,液量损失少,但是超声波产生的化学自由基团能使某些敏感性活性物质变性失活。而且大容量装置声能传递,散热均有困难,应采取相应降温措施。对超声波敏感和核酸应慎用。空化作用是细胞破坏的直接原因,同时会产生活性氧,所以要加一些巯基保护剂。展开
加trizol,之后,还需要用玻璃匀浆器磨么?
个人觉得没有必要,如果需要的话是为什么呢?是怕组织没有完全破碎么?
谢谢大家了!
本人实验新手,用玻璃匀浆器提取组织蛋白。但组织的量不是很多,加入的裂解液也不多,匀桨后裂解液包含着组织都粘在玻璃管壁上,怎么样取出我的蛋白?能离心吗,匀浆器这种玻璃管子能离心吗?
动物组织块冻存在液氮,取出后放到1.5mlEP管中,EP管中加入RIPA裂解液(按照0.1mg组织最多1ml裂解液的体积使用,用前已加入PMSF),使用电动组织研磨器研磨,每次研磨时间10秒左右,研磨过程中发现蛋白质起泡,就像做蛋糕时鸡蛋被打出泡一样。请问如何避免这种情况?
PS,本来试着在冰上操作,但是电动组织研磨器工作时EP管震动的很厉害,没法放在冰上。

