Adeno-associated Virus (AAV)
Starting Materials to Run a ddPCR for rAAV Titration with An AutoDG Droplet Generator and QX 200 Droplet Reader
Molecular Biology Grade Water, Hyclone, SH30538.02 GeneAmp 10X PCR Buffer, Applied Biosystems, N8080129 ddPCR Supermix for Probes no dUTP, Bio-Rad,1863023 10% Pluronic F-68, Thermo Fisher, 24040032 Droplet generation oil, Bio-Rad, […]
Tips and tricks for performing ddPCR
Clean everything You must have a clean working area to titer your AAV by ddPCR. Droplet digital PCR is a sensitive assay, so cross contamination into a no template control […]
Membrane Information
Polytetrafluoroethylene (PTFE)Hydrophobic membrane. Resistant to organic solvents as well as strong acids and bases. Low protein binding. Low in extractables. Main applications are the filtration of non-aqueous samples. Prior to […]
Quantification of AAV particles and empty capsids by optical density measurement
Predicted relationship of capsid to vector genome ratio (cp/vg) and absorbance ratio (A260/A280) for a denatured rAAV*. The theoretical A260/A280 ratio was plotted for the rAAV vector with a cp/vg […]
qPCR Determination of Mycoplasma in AAV Samples
Generate primers are designed to amplify the mycoplasma 16S rDNA via qPCR. Primer sequence:16S U1 (forward) 5’–GTTTGATCCTGGCTCAGGAYDAAC– 3’ **16S U8 (reverse) 5’–GAAAGGAGGTRWTCCAYCCSCAC– 3’ *** Y = C or T; D […]
AAV10 VP1 Sequence
atggctgctgacggttatcttccagattggctcgaggacaacctctctgagggcattcgcgagtggtgggacctgaaacctggagcccccaagcccaaggccaaccagcagaagcaggacgacggccggggtctggtgcttcctggctacaagtacctcggacccttcaacggactcgacaagggggagcccgtcaacgcggcggacgcagcggccctcgagcacgacaaggcctacgaccagcagctcaaagcgggtgacaatccgtacctgcggtataaccacgccgacgccgagtttcaggagcgtctgcaagaagatacgtcttttgggggcaacctcgggcgagcagtcttccaggccaagaagcgggttctcgaacctctcggtctggttgaggaagctgctaagacggctcctggaaagaagagaccggtagaaccgtcacctcagcgttcccccgactcctccacgggcatcggcaagaaaggccagcagcccgctaaaaagagactgaactttgggcagactggcgagtcagagtcagtccccgaccctcaaccaatcggagaaccaccagcaggcccctctggtctgggatctggtacaatggctgcaggcggtggcgctccaatggcagacaataacgaaggcgccgacggagtgggtagttcctcaggaaattggcattgcgattccacatggctgggcgacagagtcatcaccaccagcacccgaacctgggccctgcccacctacaacaaccacctctacaagcaaatctccaacgggacatcgggaggaagcaccaacgacaacacctacttcggctacagcaccccctgggggtattttgacttcaacagattccactgccacttctcaccacgtgactggcagcgactcatcaacaacaactggggattccggccaaaaagactcagcttcaagctcttcaacatccaggtcaaggaggtcacgcagaatgaaggcaccaagaccatcgccaataaccttaccagcacgattcaggtatttacggactcggaataccagctgccgtacgtcctcggctccgcgcaccagggctgcctgcctccgttcccggcggatgtcttcatgattccccagtacggctacctgacactgaacaatggaagtcaagccgtaggccgttcctccttctactgcctggaatattttccatctcaaatgctgcgaactggaaacaattttgaattcagctacaccttcgaggacgtgcctttccacagcagctacgcacacagccagagcttggaccgactgatgaatcctctcattgaccagtacctgtactacttatccagaactcagtccacaggaggaactcaaggtacccagcaattgttattttctcaagctgggcctgcaaacatgtcggctcaggccaagaactggctgcctggaccttgctaccggcagcagcgagtctccacgacactgtcgcaaaacaacaacagcaactttgcttggactggtgccaccaaatatcacctgaacggaagagactctctggtgaatcccggtgtcgccatggcaacccacaaggacgacgaggaacgcttcttcccgtcgagcggagtcctgatgtttggaaaacagggtgctggaagagacaatgtggactacagcagcgttatgctaacaagcgaagaagaaattaaaaccactaaccctgtagccacagaacaatacggcgtggtggctgacaacttgcagcaagccaatacagggcctattgtgggaaatgtcaacagccaaggagccttacctggcatggtctggcagaaccgagacgtgtacctgcagggtcccatctgggccaagattcctcacacggacggcaactttcacccgtctcctctgatgggcggctttggacttaaacacccgcctccacagatcctgatcaagaacacgccggtacctgcggatcctccaacaacgttcagccaggcgaaattggcttccttcatcacgcagtacagcaccggacaggtcagcgtggaaatcgagtgggagctgcagaaggagaacagcaaacgctggaacccagagattcagtacacttcaaactactacaaatctacaaatgtggactttgctgtcaatacagagggaacttattctgagcctcgccccattggtactcgttatctgacacgtaatctgtaa
Human ChAT Promoter
cccgggagcagggggtggggaaggaggagcagtggctggtggcaagtctgagccaggacatagccgatgacagcctgtgaatgaccggccagactggggttggaccttcggggcagagcaggcagcccttggaaagccaggctggtaggagcatgggtgggaggcacaaaacacagacaaacagacatatatacacatatatcctctaggcatctccgaggagtgatggaggagagagttggtaggtgggaccacaggaaaccatcccacccagctctgccttcttcaagctcagggtccctgaagcctgactagtgtctcccgaatgggggaggggggggctctgggcagagcaaaaggaggcattggctgcatgtcttagggcctttaccaactgaccttgtccttgctgtcctctcaaaagactgccatgctccggtgaatcctcaccaggacgccacctctttcactttgcagttgggggcagcccctacgaacccacagcactgggaggcgtgggaacaagcctgtggagagtgtccctgcactgtgccagcaagggctttacattctctgcccagcactgcaaggcagccccaggctcctccatccttgggcctggttcatccatctttctggactctctgttcctggcaacaaccctgccagtcccgatacatgggcggtgcttgcctctgggtaggggtggtgttgggtggggagaggggtgaggaaattagctaaacctcagctttagacttgagagaatagagggtgggctggtggaaaacatgagtatgtgcctgtggaggcttaggttcagggggatctcagagggtagcaattgtgacccacagcctaataataacagtctttgccctcttggcctggctc