TcRBP40 pull-down ribonomics

ABSTRACT: RNA pull-down assay.
For the recombinant protein pull-down assays, 50 M-5g of recombinant His-tag TcRBP40 protein were bound to 100 uL of Ni-NTA resin (Qiagen) overnight at 4M-0C. 100 M-5g of total RNA from epimastigotes were incubated with the bound protein in 500 M-5l EMSA buffer at 4M-0C for 2 h, in the presence of Heparine and Spermidine as competitors. Bounded and supernatant samples were separated. The bound sample was washed with the same buffer three times, soft-mixing for 10 min each. After washing, RNA present in the bound and supernatantM- fractions were purified.

RNA purification and amplification:

RNA was extracted using the RNeasy mini kit (Qiagen). Linearly amplified RNA (aRNA) was generated with the MessageAmpM-^YII aRNA Amplification kit (Ambion), accordin

ORGANISM(S): Trypanosoma cruzi

SUBMITTER: Christian Probst 

PROVIDER: E-MEXP-3057 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress