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PDAP1 RIP in UV-crosslinked human T cells

ABSTRACT: PDAP1 RIP in UV-crosslinked human T cells

PROVIDER: PRJNA761680 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15813454_1.fastq.gz	Fastqsanger.gz
	SRR15813454_2.fastq.gz	Fastqsanger.gz
	SRR15813455.fastq.gz	Fastqsanger.gz
	SRR15813455_1.fastq.gz	Fastqsanger.gz
	SRR15813455_2.fastq.gz	Fastqsanger.gz

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PDAP1 RIP in UV-crosslinked human T cells

Project description:Human CD4+ Total memory cells (CD4+CD25-CD45RA-CCR7+/-) were activated for 5d with anti-CD3/anti-CD28 antibodies. About 40*10^6 cells were then irradiated twice with 254nm UV light at 0.2J and then lysed in RIPA buffer. 1mg of cell extract was incubated on a rotating wheel ~16 h at 4°C with anti-PDAP1 antibody. PDAP1:mRNA complexes were immunoprecipitated with protein-G dynabeads 4hrs at 4°C and then treated with proteinase K. RNA was then purified using TRI-reagent and RNA zymospin columns. Three independent donors were used for this experiment. Inputs were also sequenced. Total RNA was sequenced in paired-end mode. This experiment is assessing direct targets bound by PDAP1 in human T cells.

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