Words similar to npm
Example sentences for: npm
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To obtain the 5' end of the NPM3 cDNA, we performed 5' Rapid Amplification of cDNA Ends (RACE), using a Marathon-Ready™ human testis cDNA library (Clontech, Palo Alto, CA) and the Marathon™ cDNA amplification kit (Clontech, Palo Alto, CA).
It will be important in future work to refine these results and test, for example, whether NPM3 may have chromatin or ribosome assembly functions that are similar or complementary to nucleoplasmin or nucleophosmin, or whether it may have completely independent functions.
We digested previously isolated lambda genomic clones of the human FGF8 region [ 22 ] with restriction endonucleases (Promega, Madison, WI) and analyzed them by Southern blotting using a mouse Npm3 cDNA [ 21 ] probe, as previously described [ 20 ] . An 8.4-kb Xho I fragment from these clones that hybridized to the probe was subcloned into pBluescript (Stratagene) for analysis.
The RACE conditions were as follows: 50 μl reactions containing 1X KlenTaq buffer and Advantage KlenTaq Polymerase Mix (Clontech, Palo Alto, CA), 0.2 mM deoxyribonucleotides, 0.2 μM NPM3 GSP1 (5'-CGG TGA GGC AGA GCA TGG TTA GTG C-3'), and 0.2 μM API primer (Clontech, Palo Alto, CA).
A cDNA that encodes a hemagglutinin-tagged NPM3 protein was produced by PCR under the following conditions: 100 μl reactions with 1X Pfu buffer, 0.2 mM deoxyribonucleotides, 1 μM HA-F primer (5'-AAA GAA TTC AGC ATG TAC CCA TAC GAC GTC CCA GAC TAC GCC GCC GCC GGT ACT GCA GCT GCC-3'), 1 μM NPM3 -R2 primer (5'-AAA GAA TTC CTA GGG CCT GCC CCC CTG CTT TTT GGC AGG AAG GAT GGG-3'), 1 ng of human NPM3 cDNA insert, and 2.5 Units of recombinant Pfu DNA Polymerase.
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