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Similarly, five out of the six mRNAs with significantly reduced levels in the puf-8 tcer-1 double mutant have been shown previously to affinity purify with PUF-8 (Mainpal et al., 2011).
Purify with a QiaQuick Nucleotide Removal kit following manufacturer's instructions.
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None of the other candidate novel RBPs has been reported to co-purify with any known RNA binding proteins; contamination with co-purifying proteins is therefore unlikely to account for these results.
Lupin flour revealed to be a difficult matrix, since large amounts of compounds tend to co-purify with DNA, even adopting well established extraction protocols.
It does not co-purify with the bacterial Hsp90 homologue, HtpG.
Swi1 is known to co-purify with Swi3 from S. pombe cell extracts [20], [24].
A potential complication of any protein purification is contamination with proteins that co-purify with the protein of interest.
This includes proteins that localize to the outer and inner mitochondrial membrane, indicating that large parts of mitochondrial membranes co-purify with dysferlin.
Spots with green signal but no red signal were kept separated as RNAs that were expressed but did not co-purify with the candidate RBP.
Recently, the human Ccr4-Not complex was purified and a number of mRNA export and nuclear pore proteins were found to co-purify with the complex [71].
While the exact mechanism for this is unclear, subunit C may play the key role as it is the only subunit that does not co-purify with either the V1 or VO domain after complex disassembly [49].
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CEO of Professional Science Editing for Scientists @ prosciediting.com