pseap control vector  (TaKaRa)

Journal: BMC Biotechnology

Article Title: Effective generation of transgenic pigs and mice by linker based sperm-mediated gene transfer.

doi: 10.1186/1472-6750-2-5

Figure Lengend Snippet: Ionic interaction analysis of mAb C and DNA. 0.3 μg of Sal I-linearized pSEAP-2 control DNA was mixed with water, medium or mAb C for 40 minutes at room temperature with occasional mixing and then subjected to 1% agarose gel electrophoresis. Lane 1: DNA in dH 2 O; lane 2 and 8: DNA in Modified Tyrode's medium without BSA (MTM); lane 3: DNA plus 0.1 μg mAb C in MTM; lane 4: DNA plus 0.3 μg mAb C in MTM; lane 5: DNA plus 1 μg mAb C in MTM; lane 6: DNA plus 3 μg mAb C in MTM; and lane 7: DNA plus 10 μg mAb C in MTM.

Article Snippet: After washing twice with centrifugation at 3000 rpm for 1.5 min, the sperm pellet was resuspended in incubation buffer containing 500 ng of 32 P-labeled pSEAP-2 control DNA (Clontech Laboratories Inc., Palo Alto, CA; catalog #6052–1, vector map available at Clontech web site) made by T 4 DNA polymerase, and incubated for another 40 min. After washing the sperm twice with incubation buffer to remove unbound 32 P-labeled DNA, samples were measured by liquid scintillation counting.

Techniques: Agarose Gel Electrophoresis, Modification

Journal: BMC Biotechnology

Article Title: Effective generation of transgenic pigs and mice by linker based sperm-mediated gene transfer.

doi: 10.1186/1472-6750-2-5

Figure Lengend Snippet: Genotype analysis of transgenic mice. Southern blot analysis of transgenic mice from F1 generation. The genomic DNA was isolated from 6-week-old mouse-tails and digested with Bgl I. The 1.3 kb Bgl I fragment of pSEAP-2 control DNA was used as a probe. The numbers in bold indicate positive detection on the blot.

Article Snippet: After washing twice with centrifugation at 3000 rpm for 1.5 min, the sperm pellet was resuspended in incubation buffer containing 500 ng of 32 P-labeled pSEAP-2 control DNA (Clontech Laboratories Inc., Palo Alto, CA; catalog #6052–1, vector map available at Clontech web site) made by T 4 DNA polymerase, and incubated for another 40 min. After washing the sperm twice with incubation buffer to remove unbound 32 P-labeled DNA, samples were measured by liquid scintillation counting.

Techniques: Transgenic Assay, Southern Blot, Isolation