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Functional Protein
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Adenovirus, AAV
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* High Titer
* Cre, FLP, ΦC31
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* Transcription Factors
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Archive for July, 2011

UPenn Vector Core Adeno-associated Virus and plasmid catalog

Lot #
Vector Name
Titer GC/ml
Map
FOR CNS APPLICATIONS
AAV Expression Vectors – by Gene
GCamP
CS0112
AAV2/1.hSynap.GCaMP3.3.SV40 (Addgene 22692 MOD)
4.73E+13

V1703
AAV2/5.hSynap.GCaMP3.3.SV40 (Addgene 22692 MOD)
2.42E+13

V2134
AAV2/9.hSynap.GCaMP3.3.SV40 (Addgene 22692 MOD)

9.28E+12

ChR2
V1779
AAV2/1.CAG.ChR2-Venus.W.SV40 (Addgene 20071)

6.16E+12

V1774
AAV2/5.CAG.ChR2-Venus.W.SV40 (Addgene 20071)

2.40E+12

V2060
AAV2/9.CAG.ChR2-Venus.W.SV40 (Addgene 20071)

5.33E+12

V1622
AAV2/9.EF1a.ChR2.YFP.hGH

3.69E+12

  
ChR2(H134R)
V2286
AAV2/1.hSyn.hChR2(H134R)-EYFP.WPRE.hGH (Addgene 26973)
9.23E+12

V2284
AAV2/9.hSyn.hChR2(H134R)-EYFP.WPRE.hGH (Addgene 26973)
3.67E+13

V2005

AAV2/1.CAG.hChR2(H134R)-mCherry.WPRE.SV40 (Addgene 20938 MOD)
3.11E+12

V1973
AAV2/5.CAG.hChR2(H134R)-mCherry.WPRE.SV40 (Addgene 20938 MOD)
2.72E+12

V1947
AAV2/9.CAG.hChR2(H134R)-mCherry.WPRE.SV40 (Addgene 20938 MOD)
6.95E+12

Cre
V2106
AAV2.CMV.PI.Cre.RBG (PennVector P1090)
8.51E+12

V1980
AAV2/1.CMV.PI.Cre.RBG (PennVector P1090)

2.16E+13

V1648
AAV2/8.CMV.PI.Cre.RBG (PennVector P1090)
2.13E+13

V2000
AAV2/9.CMV.PI.Cre.RBG (PennVector P1090)
4.92E+13

V0757
AAV2/rh10.CMV.PI.Cre.RBG (PennVector P1090)
1.13E+13

GFP-Cre
V1053

AAV2.CMV.HI.GFP-Cre.SV40 (PennVector P1415)
2.56E+12

V1673

AAV2/1.CMV.HI.GFP-Cre.SV40 (PennVector P1415)

1.18E+13

V1701

AAV2/8.CMV.HI.GFP-Cre.SV40 (PennVector P1415)
9.07E+12

V1656

AAV2/9.CMV.HI.GFP-Cre.SV40 (PennVector P1415)
6.50E+12

V2087
AAV2/9.CamKII.HI.GFP-Cre.WPRE.SV40 (PennVector P1912)
3.13E+13

  
GFP-Cre with WPRE
V1895
AAV2/9.CMV.HI.GFP-Cre.WPRE.SV40 (PennVector P2004)
9.63E+12

  
V1837
AAV2/9.hSynapsin.HI.GFP-Cre.WPRE.SV40 (PennVector P1848)
2.50E+13

  
V1803
AAV2/9.TH.HI.GFP-Cre.WPRE.SV40 (PennVector P1847)
1.06E+13

  
Lot #
Vector Name
Titer GC/ml
Map
AAV Cre-Dependent Expression Vectors
ChR2
TBD

AAV2/1.EF1.dflox.hChR2(H134R)-mCherry.WPRE.hGH (Addgene 20297)
TBD

V1449
AAV2/5.EF1.dflox.hChR2(H134R)-mCherry.WPRE.hGH (Addgene 20297)
7.93E+12

V2088
AAV2/9.EF1.dflox.hChR2(H134R)-mCherry.WPRE.hGH (Addgene 20297)
3.56E+13

V1714
AAV2/1.CAGGS.flex.ChR2.tdTomato.SV40 (Addgene 18917)
6.31E+12

V1345
AAV2/5.CAGGS.flex.ChR2.tdTomato.SV40  (Addgene 18917)
1.16E+13

V1750
AAV2/9.CAGGS.flex.ChR2.tdTomato.SV40 (Addgene 18917)
1.15E+13

V2131
AAV2/1.EF1a.DIO.hChR2(H134R)-EYFP.WPRE.hGH (Addgene 20298)
1.58E+13

V2109
AAV2/5.EF1a.DIO.hChR2(H134R)-EYFP.WPRE.hGH (Addgene 20298)
6.72E+12

V2289
AAV2/9.EF1a.DIO.hChR2(H134R)-EYFP.WPRE.hGH (Addgene 20298)
7.40E+12

NpHR
V1773
AAV2/1.EF1a.DIO.eNpHR-eYFP.WP.hGH (Addgene 20949)
4.29E+12

V1328
AAV2/5.EF1a.DIO.eNpHR-eYFP.WP.hGH (Addgene 20949)
3.71E+12

V1269
AAV2/9.EF1a.DIO.eNpHR-eYFP.WP.hGH (Addgene 20949)
3.21E+12

Arch
V1643

AAV2/1.flex.CBA.Arch-GFP.W.SV40 (Addgene 22222)

6.24E+12

V1530
AAV2/5.flex.CBA.Arch-GFP.W.SV40 (Addgene 22222)
4.73E+12

V1615
AAV2/9.flex.CBA.Arch-GFP.W.SV40 (Addgene 22222)

2.21E+12

GCamP
V2226
AAV2/1.hSynap.Flex.GCaMP3.3.SV40 (Addgene 22692 MOD)
6.00E+12

V1775
AAV2/5.hSynap.Flex.GCaMP3.3.SV40 (Addgene 22692 MOD)
9.74E+12

V2178
AAV2/9.hSynap.Flex.GCaMP3.3.SV40 (Addgene 22692 MOD)
1.26E+13

Reporter Gene
V2163
AAV2/1.CAG.FLEX.EGFP.WPRE.bGH (Allen Institute 854)
1.53E+13

  
V1676
AAV2/1.CAG.LSL.EGFP.bGH (Allen Institute 851)

4.87E+12

  
V2114
AAV2/1.CAG.FLEX.tdTomato.WPRE.bGH (Allen Institute 864)
1.02

  
V1678
AAV2/1.CAG.LSL.tdTomato.bGH (Allen Institute 856)
3.56E+12

  
V1680
AAV2/9.CAG.FLEX.EGFP.WPRE.bGH (Allen Institute 854)
4.83E+12

  
V1681
AAV2/9.CAG.LSL.EGFP.bGH (Allen Institute 851)

2.12E+12

  
V1682
AAV2/9.CAG.FLEX.tdTomato.WPRE.bGH (Allen Institute 864)
3.71E+12

  
V1683
AAV2/9.CAG.LSL.tdTomato.bGH (Allen Institute 856)
3.23E+12

  
AAV Reporter Vectors – by Promoter
Synapsin
V1480
AAV2/1.hSynapsin.EGFP.RBG
2.73E+13

  
V1479
AAV2/9.hSynapsin.EGFP.RBG
2.06E+13

  
V1481
AAV2/1.hSynapsin.TurboRFP.RBG
2.82E+13

  
V1496
AAV2/9.hSynapsin.TurboRFP.RBG
4.83E+12

  
Synapsin with WPRE
V1937
AAV2/1.hSynapsin.EGFP.WPRE.bGH (PennVector P1696)
1.58+13

  
V1857
AAV2/9.hSynapsin.EGFP.WPRE.bGH (PennVector P1696)
1.09E+13

  
V1837
AAV2/9.hSynapsin.HI.GFP-Cre.WPRE.SV40 (PennVector P1848)

2.50E+13

  
CamKII
V1706
AAV2/1.CamKII 0.4. EGFP.RBG (PennVector P1474)
9.76E+12

  
V1279
AAV2/9.CamKII 0.4. EGFP.RBG (PennVector P1474)
1.86E+13

  
V2239
AAV2/9.CaMKII.ChR2-YFP.SV40 (Stanford- Deisseroth Lab)
1.28E+13

  
V2087
AAV2/9.CamKII.HI.GFP-Cre.SV40 (PennVector P1912)
3.13E+13

  
CamKII with WPRE
V1863
AAV2/1.CamKII.EGFP.WPRE.RBG (PennVector P1917)
3.43E+13

  
V1842
AAV2/5.CamKII.EGFP.WPRE.RBG (PennVector P1917)
4.09E+13

  
V2014
AAV2/9.CamKII.EGFP.WPRE.RBG (PennVector P1917)
2.58E+12

  
TH
V1559
AAV2/1.TH.EGFP.RBG (PennVector P1646)
3.26E+13

  
V1555
AAV2/8.TH.EGFP.RBG (PennVector P1646)
1.48E+13

  
V1570
AAV2/9.TH.EGFP.RBG (PennVector P1646)
1.85E+13

  
V1904
AAV2/9.TH.ChR2-YFP.RBG (P1931)
1.83E+13

  
TH with eGFP and WPRE
V1802
AAV2/9.TH.EGFP.WPRE.RBG (PennVector P1918)
2.15E+13

  
V1803
AAV2/9.TH.HI.GFP-Cre.WPRE.SV40 (PennVector P1847)
1.06E+13

  
EF1a with eGFP and WPRE
V2052
AAV2/5.EF1a.eGFP.WPRE.RBG (P2161)
9.41E+12
1
Lot #
Vector Name
Titer GC/ml
Map
FOR CNS APPLICATIONS (cont.)
CMV
GFP reporter
V1522
AAV2.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)

2.94E+ 12

  
V2040
AAV2/1.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)
1.82E+13

  
V1255
AAV2/5JC.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)

1.13E+13

  
CS0026
AAV2/6.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)

4.14E+12

  
CS0017
AAV2/6.2.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)

5.58E+12

  
V0367
AAV2/7.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)
1.75E+13

  
CS0012
AAV2/8.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)
3.60E+13

  
CS0067
AAV2/9.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)

1.95E+13

  
TurboRFP reporter
V1371
AAV2.CMV.TurboRFP.RBG (PennVector P1471)
1.17E+13

  
V2041
AAV2/1.CMV.TurboRFP.RBG (PennVector P1471)

6.75E+12

  
V1357
AAV2/8.CMV.TurboRFP .RBG (PennVector P1471)
7.28E+12

  
V1341
AAV2/9.CMV.TurboRFP.RBG (PennVector P1471)

1.90E+13

  
Luciferase reporter
V0449
AAV2.CMV.ffluciferase.SV40 (PennVector P0105)
2.19E+13

  
V0878
AAV2/1.CMV.ffLuciferase.SV40 (PennVector P0105)
2.17E+13

  
V0838
AAV2/5.CMV.ffluciferase.SV40 (PennVector P0105)
1.46E+13

  
V0880
AAV2/8.CMV.ffLuciferase.SV40 (PennVector P0105)
1.11E+13

  
V1396
AAV2/9.CMV.ffLuciferase.SV40 (PennVector P0105)
1.56E+13

  
Beta-galactosidase reporter
V1519
AAV2.CMV.LacZ.bGH (PennVector P0102)
7.05E+12

  
V0861
AAV2/1.CMV.LacZ.bGH (PennVector P0102)
3.62E+13

  
V0607
AAV2/5.CMV.LacZ.bGH (PennVector P0102)
1.64E+13

  
V0366
AAV2/7.CMV.LacZ.bGH (PennVector P0102)
2.03E+13

  
V1019
AAV2/8.CMV.LacZ.bGH (PennVector P0102)
2.21E+13

  
V1021
AAV2/9.CMV.LacZ.bGH (PennVector P0102)
2.33E+13

  
CAGS (CB7)
GFP reporter
V1963
AAV2.CB7.CI.eGFP.RBG (PennVector P1046)
4.65E+12

  
V1828
AAV2/1.CB7.CI.eGFP.RBG (PennVector P1046)
1.93E+13

  
TBD
AAV2/5.CB7.CI.eGFP.RBG (PennVector P1046)
TBD

  
V1568
AAV2/7.CB7.CI.eGFP.RBG (PennVector P1046)
6.84E+12

  
CS0078
AAV2/8.CB7.CI.eGFP.RBG (PennVector P1046)
1.43E+13

  
CS0077
AAV2/9.CB7.CI.eGFP.RBG (PennVector P1046)
2.17E+13

  
V1885
AAV2/rh10.CB7.CI.eGFP.RBG (PennVector P1046)
9.40E+12

  
GFP reporter with WPRE
V1975
AAV2/1.CB7.CI.eGFP.WPRE.RBG (PennVector P1963)
1.99E+13

  
V2023
AAV2/5.CB7.CI.eGFP.WPRE.RBG (PennVector P1963)
2.29E+13

  
V2185
AAV2/9.CB7.CI.eGFP.WPRE.RBG (PennVector P1963)
1.99E+13

  
Other reporters
V1584
AAV2/9.CB7.CI.mCherry.RBG (PennVector P1616)
7.12E+12

  
V1376
AAV2/9.CB7.CI.TurboRFP.RBG (PennVector P1498)
4.09E+12

  
V1593
AAV2/9.CB7.CI.mCerulean.RBG (PennVector P1615)
4.27E+12

  
Other reporters with WPRE
V2074
AAV2/1.CB7.CI.mCherry.WPRE.RBG (PennVector P1969)
5.26E+12

  
V2299
AAV2/9.CB7.CI.mCherry.WPRE.RBG (PennVector P1969)
7.01E+12

  
V2162
AAV2/1.CB7.CI.TurboRFP.WPRE.RBG

1.01E+13

  
V2290
AAV2/9.CB7.CI.TurboRFP.WPRE.RBG

9.24E+12

  
V2091
AAV2/1.CB7.CI.mCerulean.WPRE.RBG (PennVector P1970)
9.56E+12

  
V2309
AAV2/9.CB7.CI.mCerulean.WPRE.RBG (PennVector P1970)
TBD

  
AAV Vectors for Gene Silencing
V0623
AAV2/9.U6.shRLuc.CMV.ZsGreen.SV40 (PennVector P0111)
1.06E+13

  
 
back to top

    
Lot #
Vector Name
Titer GC/ml
Map
FOR LIVER APPLICATIONS
AAV Expression Vectors – by Promoter
TBG (liver-specific)
CS0119
AAV2/8.TBG.PI.Cre.RBG (PennVector P1091)

3.71E+13

  
V1693
AAV2/8.TBG.PI.EGFP.bGH (PennVector P1014)
1.99E+13

  
TBD
AAV2/8.TBG.nLacZ.bGH (PennVector P0141)
TBD

  
V1811
AAV2/8.TBG.LacZ.bGH (PennVector P0142)
2.63E+13

  
V2140
AAV2/8.TBG.PI.ffluciferase.RBG

4.62E+12

  
V0497
AAV2/9.TBG.PI.EGFP (PennVector P1014)
5.04E+13

  
CAGS (CB7)
GFP Reporter
CS0078
AAV2/8.CB7.CI.EGFP.RBG (PennVector P1046)
1.43E+13

  
CS0077
AAV2/9.CB7.CI.EGFP.RBG (PennVector P1046)
2.17E+13

  
V1975
AAV2/1.CB7.CI.EGFP.WPRE.RBG (PennVector P1963)
1.99E+13

  
V2185
AAV2/9.CB7.CI.EGFP.WPRE.RBG (PennVector P1963)
1.99E+13

  
Other reporters
V1584
AAV2/9.CB7.CI.mCherry.RBG (PennVector P1616)
7.12E+12

  
V1376
AAV2/9.CB7.CI.TurboRFP.RBG (PennVector P1498)
4.09E+12

  
V1593
AAV2/9.CB7.CI.mCerulean.RBG (PennVector P1615)
4.27E+12

  
Other reporters with WPRE
V2299
AAV2/9.CB7.CI.mCherry.WPRE.RBG (PennVector P1969)
7.01E+12

  
V2290
AAV2/9.CB7.CI.TurboRFP.WPRE.RBG

9.24E+12

  
V2309
AAV2/9.CB7.CI.mCerulean.WPRE.RBG (PennVector P1970)
TBD

  
AAV Vectors for Gene Silencing
V0623
AAV2/9.U6.shRLuc.CMV.ZsGreen.SV40 (PennVector P0111)
1.06E+13

  
 
back to top

    
Lot #
Vector Name
Titer GC/ml
Map
FOR HEART APPLICATIONS
AAV Expression Vectors – by Promoter
TNT (heart specific)
V1978
AAV2/9.cTnT.PI.EGFP.WPRE.RBG (PennVector P1967)
2.10E+13

  
CMV
GFP reporter
V1522
AAV2.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)

2.94E+12

  
CS0026
AAV2/6.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)
4.14E+12

  
CS0017
AAV2/6.2.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)
5.58E+12

  
CS0067
AAV2/9.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)
1.95E+13

  
TurboRFP reporter
V2041
AAV2/1.CMV.TurboRFP.RBG (PennVector P1471)
6.75E+12

  
V1341
AAV2/9.CMV.TurboRFP.RBG (PennVector P1471)

1.90E+13

  
Luciferase reporter
V0878
AAV2/1.CMV.ffLuciferase.SV40 (PennVector P0105)
2.17E+13

  
V1396
AAV2/9.CMV.ffLuciferase.SV40 (PennVector P0105)
1.56E+13

  
Beta-galactosidase reporter
V0861
AAV2/1.CMV.LacZ.bGH (PennVector P0102)
3.62E+13

  
V1021
AAV2/9.CMV.LacZ.bGH (PennVector P0102)
2.33E+13

  
Cre
V1980
AAV2/1.CMV.PI.Cre.RBG (PennVector P1090)

2.17E+13

V2000
AAV2/9.CMV.PI.Cre.RBG (PennVector P1090)

4.92E+13

Cre-GFP Reporter
V1673
AAV2/1.CMV.HI.GFP-Cre.SV40 (PennVector P1415)
1.18E+13

  
V1656
AAV2/9.CMV.HI.GFP-Cre.SV40 (PennVector P1415)
6.50E+12

  
CAGS (CB7)
GFP Reporter
V1828
AAV2/1.CB7.CI.EGFP.RBG (PennVector P1046)
1.93+13

  
CS0077
AAV2/9.CB7.CI.EGFP.RBG (PennVector P1046)
2.17E+13

  
V1975
AAV2/1.CB7.CI.EGFP.WPRE.RBG (PennVector P1963)
1.99E+13

  
V2185
AAV2/9.CB7.CI.EGFP.WPRE.RBG (PennVector P1963)
1.99E+13

  
Other reporters
V1584
AAV2/9.CB7.CI.mCherry.RBG (PennVector P1616)
7.12E+12

  
V1376
AAV2/9.CB7.CI.TurboRFP.RBG (PennVector P1498)
4.09E+12

  
V1593
AAV2/9.CB7.CI.mCerulean.RBG (PennVector P1615)
4.27E+12

  
AAV Vectors for Gene Silencing
V0623
AAV2/9.U6.shRLuc.CMV.ZsGreen.SV40 (PennVector P0111)
1.06E+13

  
 
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Lot #
Vector Name
Titer GC/ml

  
FOR MUSCLE APPLICATIONS
AAV Expression Vectors – by Promoter
MCK6 (muscle-specific)
V1477
AAV2/9.MCK6.EGFP.RBG (PennVector P1607)
2.12E+13

  
CMV
GFP reporter
V2040
AAV2/1.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)

1.82E+13

  
CS0012
AAV2/8.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)
3.60E+13

  
CS0067
AAV2/9.CMV.PI.EGFP.WPRE.bGH (PennVector P0101)
1.95E+13

  
TurboRFP reporter
V2041
AAV2/1.CMV.TurboRFP.RBG (PennVector P1471)

6.75E+12

  
V1357
AAV2/8.CMV.TurboRFP .RBG (PennVector P1471)
7.28E+12

  
V1341
AAV2/9.CMV.TurboRFP.RBG (PennVector P1471)
1.90E+13

  
Luciferase reporter
V0878
AAV2/1.CMV.ffLuciferase.SV40 (PennVector P0105)
2.17E+13

  
V0880
AAV2/8.CMV.ffLuciferase.SV40 (PennVector P0105)
1.11E+13

  
V1396
AAV2/9.CMV.ffLuciferase.SV40 (PennVector P0105)
1.56E+13

  
Beta-galactosidase reporter
V0861
AAV2/1.CMV.LacZ.bGH (PennVector P0102)
3.62E+13

  
V1019
AAV2/8.CMV.LacZ.bGH (PennVector P0102)
2.21E+13

  
V1021
AAV2/9.CMV.LacZ.bGH (PennVector P0102)
2.33E+13

  
Cre
V1980
AAV2/1.CMV.PI.Cre.RBG (PennVector P1090)
2.17E+13

V1648
AAV2/8.CMV.PI.Cre.RBG (PennVector P1090)
2.13E+13

V2000
AAV2/9.CMV.PI.Cre.RBG (PennVector P1090)
4.92E+13

GFP-Cre
V1673
AAV2/1.CMV.HI.GFP-Cre.SV40 (PennVector P1415)
1.18E+13

  
V1701
AAAV2/8.CMV.HI.GFP-Cre.SV40 (PennVector P1415)
9.07E+12

  
V1656
AAV2/9.CMV.HI.GFP-Cre.SV40 (PennVector P1415)
6.50E+12

  
CAGS (CB7)
GFP reporter
V1828
AAV2/1.CB7.CI.EGFP.RBG (PennVector P1046)
1.93E+13

  
CS0078
AAV2/8.CB7.CI.EGFP.RBG (PennVector P1046)
1.43E+13

  
CS0077
AAV2/9.CB7.CI.EGFP.RBG (PennVector P1046)
2.17E+13

  
V1975 AAV2/1.CB7.CI.EGFP.WPRE.RBG (PennVector P1963) 1.99E+13  
V2185 AAV2/9.CB7.CI.EGFP.WPRE.RBG (PennVector P1963)
1.99E+13

  
Other reporters
V1584
AAV2/9.CB7.CI.mCherry.RBG (PennVector P1616)
7.12E+12

  
V1376
AAV2/9.CB7.CI.TurboRFP.RBG (PennVector P1498)
4.09E+12

  
V1593
AAV2/9.CB7.CI.mCerulean.RBG (PennVector P1615)
4.27E+12

  
AAV Vectors for Gene Silencing
V0623
AAV2/9.U6.shRLuc.CMV.ZsGreen.SV40 (PennVector P0111)
1.06E+13

  
 
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Lot #
Vector Name
Titer GC/ml

  
FOR LUNG APPLICATIONS
CC10 (clara cell/lung specific)
TBD
AAV2/9.CC10.TurboRFP
TBD

  
CAGS (CB7)
GFP reporter
V1828
AAV2/1.CB7.CI.EGFP.RBG (PennVector P1046)
1.93E+13

  
V1841
AAV2/5.CB7.CI.EGFP.RBG (PennVector P1046)
1.68E+13

  
CS0077
AAV2/9.CB7.CI.EGFP.RBG (PennVector P1046)
2.17E+13

  
V1885
AAV2/rh10.CB7.CI.EGFP.RBG (PennVector P1046)
9.40E+12

  
V1975
AAV2/1.CB7.CI.EGFP.WPRE.RBG (PennVector P1963)
1.99E+13

  
V2185
AAV2/9.CB7.CI.EGFP.WPRE.RBG (PennVector P1963)
1.99E+13

  
Other reporters
V1584
AAV2/9.CB7.CI.mCherry.RBG (PennVector P1616)
7.12E+12

  
V1376
AAV2/9.CB7.CI.TurboRFP.RBG (PennVector P1498)
4.09E+12

  
V1593
AAV2/9.CB7.CI.mCerulean.RBG (PennVector P1615)
4.27E+12

  
AAV Vectors for Gene Silencing
V0623
AAV2/9.U6.shRLuc.CMV.ZsGreen.SV40 (PennVector P0111)
1.06E+13

ALL ADDGENE PLASMIDS MUST BE OBTAINED FROM THE ADDGENE WEBSITE

AAV AD HELPER PLASMID
stock
pAd DeltaF6
stock

AAV REP CAP PLASMIDS (TRANS PLASMIDS)
P0001
pAAV2/1 (PennVector P0001)
P0001
P0002
pAAV2 (PennVector P0002)
P0002
P0003
pAAV2/5 (PennVector P0003)
P0003
P0004
pAAV2/6 (PennVector P0004)
P0004
P0005
pAAV2/6.2 (PennVector P0005)
P0005
P0006
pAAV2/7 (PennVector P0006)
P0006
P0007
pAAV2/8 (PennVector P0007)
P0007
P0008
pAAV2/9 (PennVector P0008)
P0008
P0009
pAAV2/rh10 (PennVector P0009)
P0009
AAV EXPRESSION PLASMIDS FOR INSERTION (CIS PLASMIDS)
Parental Plasmids
P0100
pZac2.1 aka pAAV.CMV.PI.SV40 (PennVector P0100)
P0100
TBD
pENN.AAV.CMV.PI.RBG (PennVector TBD)
TBD
P1044
pENN.AAV.CB7.CI.RBG (PennVector P1044)
P1044
P1015
pENN.AAV.TBG.PI.RBG (PennVector P1015)
P1015
AAV EXPRESSION PLASMIDS FOR REPLACEMENT (CIS PLASMIDS)
Reporter Plasmids
P1060
pENN.AAV.CMV.PI.EGFP.RBG (PennVector P1060)
P1060
P1046
pENN.AAV.CB7.CI.EGFP.RBG (PennVector P1046)
P1046
P1963
pENN.AAV.CB7.CI.EGFP.WPRE.RBG (PennVector P1963)
P1963
P1014
pENN.AAV.TBG.PI.EGFP.RBG (PennVector P1014)
P1014
P1372
pENN.AAV.cTNT.PI.EGFP.RBG (PennVector P1372)
P1372
P1967
pENN.AAV.cTNT.PI.EGFP.WPRE.RBG (PennVector P1967)
P1967
P1626
pENN.AAV.hSynapsin.EGFP.RBG (PennVector P1626)
P1626
P1696
pAAV.hSynapsin.EGFP.WPRE.RBG (PennVector P1696)
P1696
P1474
pENN.AAV.CamKII 0.4. EGFP.RBG (PennVector P1474)
P1474
P1917
pENN.AAV.CamKII 0.4. EGFP.PRE.RBG (PennVector P1917)
P1917
P1646
pENN.AAV.TH.EGFP.RBG (PennVector P1646)
P1646
P1918
pENN.AAV.TH.EGFP.WPRE.RBG (PennVector P1918)
P1918
SIRNA PLASMIDS FOR CLONING (REPLACEMENT)
Reporter Plasmids
P0111
pAAV.U6.shRLuc.CMV.ZsGreen.SV40 (PennVector P0111)
P0111
P1867
pAAV.U6.shRLuc.CMV.EGFP.SV40 (PennVector P1867)
P1867
P1781
pAAV.H1.shRLuc.CMV.ZsGreen.SV40 (PennVector P1781)
P1781
P1782
pAAV.H1.shRLuc.CMV.TurboRFP.SV40 (PennVector P1782)
P1782
P1537
pAAV.shR(Target).CB7.CI.TurboRFP.SV40 (PennVector P1537)
P1537
Lot #

LENTIVIRAL PLASMID NAME
Vector NTI
P1324

pLenti6.2GW.SIN.cPPT.WP (modified from Invitrogen) (PennVectorP1324)
P1324
Lot #

ADENOVIRAL & SHUTTLE PLASMID NAME
Vector NTI
P0670
pH5′.040.pkGFP-II (PennVector P0670)
P0670
P0503
pShuttle (Clontech) (PennVectorP0503)
P0503

Comments

July 31, 2011 at 6:38 pm ·

How to set up wwwblast on Ubuntu and other linux distributions

1. cd to your web directory

cd /home/myweb/webapps

Download wwwblast from ftp://ftp.ncbi.nlm.nih.gov/blast/executables/release/LATEST, e.g,

wget ftp://ftp.ncbi.nlm.nih.gov/blast/executables/release/LATEST/wwwblast-2.2.25-ia32-linux.tar.gz

2. extract file

tar xzf wwwblast-2.2.25-ia32-linux.tar.gz

3. Change index.html to another file name

mv index.html index_old.html
mv blast_cs.html index.html

4. Modifying the apache configuration file
create a new file /etc/httpd/conf.d/blast.conf

vi /etc/httpd/conf.d/blast.conf

5. Add the following lines to the blast.conf file

# for wwwblast
AddHandler cgi-script .cgi
<directory “/home/myweb/webapps/blast”>
Options FollowSymLinks +ExecCGI +Indexes
</directory>

6. Restart apache

sudo apachectl graceful

7. Use this sequence to test blast

TACTGTTATCGATCCGGTCGAAAAACTGCTGGCAGTGGGGCATTACCTCGAATCTACCGTCGATATTGCT

8. Make changes to your index.html

cp index.html index_long.html
vi index.html

9. use the following lines for your index.html file
<HTML>
<TITLE>BLAST Search </TITLE>
<BODY BGCOLOR="#FFFFFF" LINK="#0000FF" VLINK="#660099" ALINK="#660099">
<FORM ACTION="blast_cs.cgi" METHOD = POST NAME="MainBlastForm" ENCTYPE= "multipart/form-data">
<B>Choose program to use and database to search:</B>
<P>
<a href="docs/blast_program.html">Program</a>
<select name = "PROGRAM">
    <option> blastn
    <option> blastp
    <option> blastx
    <option> tblastn
    <option> tblastx
</select>
<a href="docs/blast_databases.html">Database</a>
<select name = "DATALIB">
    <option VALUE = "test_na_db"> test_na_db
    <option VALUE = "test_aa_db"> test_aa_db
</select>
<P>
Enter here your input data as
<select name = "INPUT_TYPE">
    <option> Sequence in FASTA format
    <option> Accession or GI
</select>
<BR>
<textarea name="SEQUENCE" rows=6 cols=60>
</textarea>
<BR>
<INPUT hidden TYPE="checkbox" NAME="OVERVIEW"  CHECKED>
<INPUT TYPE="button" VALUE="Clear sequence" onClick="MainBlastForm.SEQUENCE.value=”;MainBlastForm.QUERY_FROM.value=”;MainBlastForm.QUERY_TO.value=”;MainBlastForm.SEQUENCE.focus();">
<INPUT TYPE="submit" VALUE="Search">
</FORM>
<HR>
</BODY>
</HTML>

Comments

July 29, 2011 at 7:24 am ·

BL21 cells for toxic protein expression

1, BL21 Star (DE3)pLysS, F- ompT hsdSB (rB-mB-) gal dcm rne131 (DE3) pLysS (CamR), from Invitrogen, Cat #: C6565-03

The pLysS plasmid carried by the BL21 StarTM(DE3)pLysS strain produces T7 lysozyme to reduce basal level expression of the gene of interest. pLysS confers resistance to chloramphenicol (CamR) and contains the p15A origin. This origin allows pLysS to be compatible with pUC- or pBR322-derived plasmids. It requires IPTG to induce expression of the T7 RNA polymerase from the lacUV5 promoter.

RNaseE (rne131), which is one of the major sources of mRNA degradation. BL21 Star cells significantly improve the stability of mRNA transcripts and increase protein expression yield from T7 promoter-based vectors due to rne131 mutation.

2. Rosetta(DE3)pLysS. Novagen, EMD Biosciences

F- ompT hsdSB(rB- mB-) gal dcm (DE3) pLysSpRARE2 (Cam-R)

Rosettacells have tRNAs for the codons AUA, AGG, AGA, CUA, CCC, and GGA on a compatible chloramphenicol-resistant plasmid, pRARE. pLysS is a chloramphenicol-resistant plasmid with a P15A replicon that encodes T7 lysozyme, which is for inhibiting T7 RNA polymerase. pLysSRARE and pLysSRARE2 contain a CamR plasmid (pACYC184) that carries the gene for T7 lysozyme plus tRNA genes for several codons rarely used in E. coli.

3. Lemo21(DE3), NEB. Cat # C2528H.

fhuA2 [lon] ompT gal (λ DE3) [dcm] ∆hsdS/ pLemo(CamR)
λ DE3 = λ sBamHIo ∆EcoRI-B int::(lacI::PlacUV5::T7 gene1) i21 ∆nin5
pLemo = pACYC184-PrhaBAD-lysY

Lemo21(DE3) offers the host features of BL21(DE3) while also allowing for tunable expression of difficult clones. Tunable expression is achieved by varying the level of lysozyme (lysY), the natural inhibitor of T7 RNA polymerase. The level of lysozyme is modulated by adding L-rhamnose to the expression culture at levels from zero to 2000 µM. When Lemo21(DE3) is grown without rhamnose, the strain performs the same as a pLysS containing strain. However, optional addition of rhamnose tunes the expression of the protein of interest. For difficult soluble proteins, tuning the expression level may also result in more soluble, properly folded protein.

4. OverExpress C41(DE3) and C43(DE3), Lucigen, 60448-1, 60446-1

C41(DE3) was derived from BL21(DE3). This strain has at least one mutation, which prevents cell death associated with expression of many recombinant toxic proteins. The strain C43(DE3) was derived from C41(DE3) by selecting for resistance to a different toxic protein and can express a different set of toxic proteins to C41(DE3).

Comments

July 26, 2011 at 5:02 pm ·

Purification of adeno-associated virus (AAV) by FPLC using heparin affinity column

1. Equilibrate HiTrap heparin affinity column with 10 ml PBS-MK at 1 ml/min.

2. Load the column with AAV at 0.3 ml/min.

3. Collect the flow-through in a 14-ml tube.

4. Wash column with 5 ml of PBS-MK buffer using a syringe pump at the rate of 0.3 ml/min.

5. Collect the flow-through in a 14-ml tube.

6. Elute AAV from the column with 2.4 ml of 1 M NaCl/PBS-MK buffer.

7. Collect the first 0.4 ml in a microcentrifuge tube. This is the ‘void volume’.

8. After this initial 0.4-ml void volume, collect the following fractions: fraction 1 is 1 ml, fraction 2 is 0.5 ml and fraction 3 is 0.5 ml. The AAV should come off the column in the first 1.5 ml (fractions 1 and 2).

9. Desalt using a HiTrap desalting column. Wash the HiTrap desalting column with 25 ml of PBS in a 30-ml syringe.

Do not exceed a flow rate of 5 ml/min.

10. Prepare the following:
A 3-ml syringe containing 1.5 ml of fractions 1 and 2.
A 3-ml syringe containing 3 ml of PBS.

11. Position the column to collect fractions into microcentrifuge tubes: the first tube for the 1.5 ml of void flow-through, then ‘exchange’ fractions X1–X3 (1 ml each).

12. Load the 1.5 ml of AAV into the 3-ml syringe.

13. Quickly load the 3 ml of PBS in the second 3-ml syringe.

14. The AAV should be in fractions X1 and X2. Pool fractions X1 and X2 and make 10 aliquots of 200 μl each (or other aliquot sizes) in microcentrifuge tubes. Prepare 1:10 and 1:100 dilutions of the AAV stock in TE buffer for determining titer later.

The vector stock and dilutions can be stored at –80 °C indefinitely. If necessary, store stocks in small aliquots and avoid repeated freezing and thawing.

Comments

July 25, 2011 at 9:49 am ·

Protocol for Adeno-associated virus (AAV) production using GeneExpresso Max

1. Thaw a vial of 293T cells (each vial contains ~3 × 10^6 cells) and plate cells in a 10-cm dish containing 10 ml D10 (DMEM with 10% FBS). Maintain in a 37 °C incubator with 5% CO2 atmosphere.

2. After 3 days, aspirate the medium and wash the dish with 2 ml of 1× PBS. Add 2 ml of trypsin-EDTA solution and incubate at 37 °C for 2 min or until cells detach from culture dish.

3. Add 4 ml of D10 to neutralize trypsin-EDTA solution and transfer the cell suspension to a 15-ml conical tube.

4. Centrifuge at 250g for 5 min at 20 °C in a tabletop centrifuge.

5. Resuspend the cells in 1 ml of D10. Take a 10-μl aliquot and count the cells using a hemocytometer.

6. Adjust cell concentration to 10^6 cells per ml with D10 and plate 1 ml of cell suspension in eight 10-cm dishes, each contains 10 ml D10. Culture for 72 h in a 37 °C incubator with 5% CO2 atmosphere.

7. After the 3-d incubation, harvest cells as described in Steps 2–4. Resuspend in 5 ml of D10 and take a 10-μl aliquot for counting with a hemocytometer. Adjust cell concentration to 4 × 10^6 cells per ml and plate 1 ml of cell suspension in 30 10-cm dishes, each containing 10 ml D10. Incubate in a 37 °C incubator overnight.

8. The next day, prepare solution A by combining 7.5 ml of DMEM (without serum) and 1 ml GeneExpresso Max in a 50-ml tube and, mixing well.

9. In another 50-ml tube, prepare solution B by combining 7.5 ml of DMEM (without serum) and 350 μg of plasmid.

10. Add solution A to solution B, mix well and incubate for 20 min. Add 0.5 ml of the transfection mix to each of the 30 dishes of 293T cells.

11. Agitate the dishes gently to evenly distribute the precipitate and return the dishes to the 37 °C incubator.

12. After 48 h, prepare dry ice/ethanol bath by placing a centrifuge tube rack in a styrofoam container. Add ~2 lbs of dry ice, pour 2 liters of ethanol over it and cover the container.

13. Collect the cells and medium by scraping the cells off the dish with a rubber policeman and transferring it to a 50-ml conical tube. Use Corning 50-ml conical tubes, as they are more resistant to the subsequent freeze-thaw cycles. Rinse dishes with 5 ml of 1× PBS and transfer it to the same conical tube. Harvest four dishes at a time into the same 50-ml tube.

14. Centrifuge at 250g for 5 min at 20 °C in a tabletop centrifuge.

15. Aspirate the medium from the conical tube and repeat Steps 14–15 until cells are pelleted from all 30 dishes. The same tube can be used for pelleting cells from all the dishes.

16. Resuspend the final pellet from the 30 10-cm dishes in 8 ml of cell lysis buffer.

17. Freeze the pellet in the dry ice/ethanol bath and thaw in a 37 °C water bath.

18. Repeat Step 18 twice and store at − 80 °C.

The pellet can be stored at − 80 °C indefinitely and thawed at your convenience.

19. Thaw cell lysate in a 37 °C water bath.

20. Add Benzonase to the cell lysate at a final concentration of 50 U ml − 1. Benzonase is added to eliminate cellular DNAs/RNAs as well as excess plasmid DNAs present during transfection.

21. Incubate at 37 °C in a water bath for 30 min. Prepare iodixanol gradient during the incubation.

22. Centrifuge at 5,000g for 30 min at 4 °C in a Sorvall high-speed centrifuge using an HS-4 rotor.

23. Collect vector-containing supernatant. The volume of the supernatant is approximately 5 ml.

24. Load the 5 ml of vector-containing supernatant over the iodixanol density gradient prepared in Step 22. Top off the tube with cell lysis buffer. This density gradient purification step can remove contaminants that may influence transduction and targeting frequencies.

25. Centrifuge at 461,300g for 1 h at 18 °C with maximum acceleration and deceleration in a Beckman Ti70 rotor on a Beckman ultracentrifuge; use proper spacers for tubes.

26. Puncture the tube on the side slightly below (3–5 mm) the 60–40% interface with an 18-gauge needle (bevel up) attached to a 10 ml syringe.

27. Collect 3–4 ml from each centrifuge tube by aspiration using the same needle. Avoid the proteinaceous material near the 40–25% interface.

28. To use the vector without removing iodixanol, proceed directly with Step 30; iodixanol is inert and non-toxic to cells. Alternatively, further purify AAV vector stocks over a HiTrap heparin affinity column, followed by desalting over a HiTrap desalting column.

Comments

July 25, 2011 at 9:40 am ·

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