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INTEGRATEDDNATECHNOLOGIESPRODUCTSFUNCTIONAL GENOMICSEndogenous Gene Positive Control Duplexes and Primers (continued)IDT recommends studying functional transfection efficiency by examining mRNA levels at 24 hours post-transfection.Alternatively, Western blots can be performed at 48-72 hours. Validated primers for HPRT SYBR® Green qRT-PCR assays areavailable for the following organisms:SYBR®-Green Q-PCR Primer SetsHPRT SYBR®-Green primers (Human) - 1 nmole or 5 nmolesHPRT SYBR®-Green primers (Mouse) - 1 nmole or 5 nmolesHPRT SYBR®-Green primers (Rat) - 1 nmole or 5 nmolesHPRT SYBR®-Green primers (Chinese hamster) - 1 nmole or 5 nmolesHPRT SYBR®-Green primers (Cow) - 1 nmole or 5 nmolesHPRT SYBR®-Green primers (Pig) - 1 nmole or 5 nmolesIt is often useful to have cloned gene fragments available to establish quantitative standard curves when performing qRT-PCR.Relative RNA levels can be measured without use of copy number standards (ΔΔCt method); however, cloned copy numberstandards permit true quantitative measurements to be made. The assay amplicon for each of the above HPRT positive controlqRT-PCR reactions has been cloned into pBlueScript TM -II and sequence verified. Clones are provided as 0.5 µg of purified,linearized plasmid DNA that is directly ready for use in PCR.Cloned Purified PlasmidsHPRT Cloned qControl (Human) - 0.5 µgHPRT Cloned qControl (Mouse) - 0.5 µgHPRT Cloned qControl (Rat) - 0.5 µgHPRT Cloned qControl (Chinese hamster) - 0.5 µgHPRT Cloned qControl (Cow) - 0.5 µgHPRT Cloned qControl (Pig) - 0.5 µgAccession#NM_000194NM_013556NM_012583J00060NM_001034035NM_001032376SYBR®-Green is a registered trademark of Invitrogen.©2008 Integrated DNA Technologies 20 www.idtdna.com