Protein sequences of Arabidopsis thaliana ASO, QS, QPT, NMNAT, NS (listed in Table 1), human NAMPT (NP_005737), yeast SIR2 (NP_010242), and human SIRT4 (NP_036372) were used in TBLASTN against JGI (Joint Genome Institute) Chlamydomonas reinhardtii genome version 4.0 (JGI v4.0, -psf.org/Chlre4/Chlre4.home.html) with expected E-values less than or equal to 1E-5 (1E-3 for SIR2 and SIRT4). The resultant genes were checked for EST coverage. Genes without full-length EST coverage, QS, NMNAT, NS, NAMPT, SRT1, and SRT2, were subjected to exon-intron predictions using GreenGenie 2 ( -bin/greengenie2/greenGenie2) [33]. The predicted coding regions were used as guidelines in primer design for RT-PCR to amplify the actual coding regions of these genes.
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Chlamydomonas BAC DNA was prepared using Qiagen Plasmid Midiprep kit. To prepare the pNIC15a plasmid, the BAC (32L22) DNA was digested with XmaI and a 6.1 kb fragment was isolated and cloned into a pBlueScript II SK vector (Stratagene, La Jolla, CA). This fragment contains a 1 kb upstream sequence, the full-length NIC15 gene, and a 2.5 kb downstream sequence, which is predicted to be part of an unknown zinc finger protein (protein id 150664). To prepare the pNIC1-56 plasmid, a 7.1 kb KpnI fragment from the BAC (10M24) DNA was cloned into a pBlueScript II SK vector. The plasmid contains a 0.7 kb upstream sequence, the full-length NIC1 gene, and a 4.7 kb downstream sequence, which is predicted to contain an unknown protein that has a HAD-superfamily hydrolase domain.
I'm trying to figure out what PP skills I need to develop now before going to b-school. I plan on applying to b-school in the fall and gun for consulting jobs but want to start practicing now before school since I am not a consultant. I heard advanced PP skills are needed similar to Excel skills for an IB associate. How advanced do I need to be to successfully complete tasks as an associate at MBB/2nd tier consulting firms? Are there any good tutorials out there or can I just learn on the job? Thanks. 2ff7e9595c
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