Download Cancer Genomics and Proteomics: Methods and Protocols by Narendra Wajapeyee PDF

By Narendra Wajapeyee

Cancer Genomics and Proteomics: equipment and Protocols, moment Edition contains tools for the analyses of melanoma genome and proteome that experience illuminated us concerning the alterations in melanoma cells. Written within the hugely winning Methods in Molecular Biology series structure, chapters contain introductions to their respective issues, lists of the required fabrics and reagents, step by step, with no trouble reproducible laboratory protocols, and key tips about troubleshooting and warding off recognized pitfalls.

Authoritative and functional, Cancer Genomics and Proteomics: equipment and Protocols, moment Edition seeks to help scientists within the extra learn into numerous elements of tumor initiation and progression.

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Wait for 5 min, and then remove the remaining ethanol. Dry the beads for 10 min in a fume hood. 5. Add 20 μl of EB buffer, then seal, vortex, and spin down. 5 Multiplexed Adapter Ligation 1. The multiplexed adapter ligation reaction is carried out in a 30 μl reaction. A master mix is prepared according to the number of samples based on the following reaction setup (Table 3). 2. Add 20 μl of master mix to each of the well containing the SPRI beads. 28 Arvindhan Nagarajan et al. 5 U/μl) 10 U DNA templates Final volume ( μl) 20 2 24 4 24 H2O 126 Total 200 3.

Mix, spin at maximum speed for 5 min, and transfer supernatant to a fresh tube. Precipitate DNA with ethanol and glycogen, and wash pellet with 70 % ethanol. 2. 5× TBE gel without ethidium bromide. Add 1 μl (50 ng) carrier DNA to each sample. Load DNA samples into the gel wells with sufficient space between adjacent samples along with marker. Excise the DNA ladder lanes from the gel and stain with SYBR Green I. With the DNA ladder as the guide cut out gel between 160 and 340 bp (these sizes correspond to 40 and 220 bp restriction fragments, as they run slower than the marker).

8. Glycerol (cell culture grade). 9. Plasmid Plus Maxi kit. 10. 293T cells. 11. U2OS cells. 12. DMEM media. 13. McCoy’s 5A media. 14. Opti-MEM media. 15. Fetal bovine serum (FBS). 16. Penicillin/streptomycin supplement. 17. TransIT-293 transfection reagent. 18. 8 mg/mL Polybrene, cell culture grade. 19. 1 mg/mL Puromycin, cell culture grade. Synthetic Lethal RNAi Screen 47 20. TSC2 antibody. 21. Hot-start Taq polymerase. 22. DNA gel purification kit. 2 Equipment 1. Electroporator (for electroporating competent bacteria).

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