By N.J. Clifton
Read Online or Download Cardiac Cell and Gene Transfer: Principles, Protocols, and Applications (Methods in Molecular Biology Vol 219) PDF
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Soils into which crop vegetation root and from which they receive crucial minerals and water include large arrays of microbes. Many have hugely worthy results on crop development and productiveness, others are pathogens inflicting illnesses and losses to yield and caliber, a couple of microbes provide safety from those pathogenic varieties and others have very little influence.
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Additional resources for Cardiac Cell and Gene Transfer: Principles, Protocols, and Applications (Methods in Molecular Biology Vol 219)
71, 3293–3298. , Hartigan-O’Connor, D. , Scott, J. , and Chamberlain, J. S. (2002) Packaging cell lines for gutted adenoviral vector growth using E1, E2b, and E3-deleted helper viruses. In press. Graham, F. , Russell, W. , and Nairn, R. (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J. Gen. Virol. 36, 59–72. , Begy, C. , and Chamberlain, J. S. (1996) Improved adenovirus packaging cell lines to support the growth of replication-defective gene-delivery vectors.
3). An SV40 polyA sequence is included in this plasmid to facilitate the expression of the trans-spliced gene product. The KpnI/SnaBI sites in pDD295 are designed for introducing the splicing acceptor signal. 4. PCR is used to introduce the splicing donor sequence into the pre-Donor vector. Donor vector is composed of a 20–25-mer oligonucleotide located upstream of a unique restriction site in the cDNA used for cloning the final product. The design of this primer should follow the general PCR primer designing principles.
3. Helper plasmid for type 1 and type 5 AAV: p5E18(2/1) for rAAV-1 packaging (33). pAV2-Rep and pAV5-Trans for rAAV-5 packaging (34). These plasmids provide viral replication and structural proteins for pseudo-packaging a type 2 rAAV genome into type 1 and type 5 capsids (see Note 7). 4. 5 M CaCl2. Sterilize by filtration and store at –20°C. 5. 05. Sterilize by filtration and store at –20°C. 05 in order to achieve high transfection efficiency. 6. DNAse I (Sigma D4513, 11 mg protein/vial, total 33 K [kuniz] units) (see Note 8).