Synthesis of HIVl RNA Standard

For quantitation，a full-length HIV1 RNA was synthesized in vitro by T7 RNA polymerase（Stratagene，La Jolla，CA）using pBKBH1OS DNA as a template.The template pBKBH1OS，kindly provided by Dr.John Rossi（NIH，Bethesda，MD），contains full-length genomec HIV1（9 kb）less the LTR cloned downstream of the T7 promoter of plasmid pBluescript II-KS（+）（Stratagene）.The size and integrity of synthesized HIV1 cRNA was confirmed by Northern analysis，and HIV1 cRNA was quantitated by OD206 measurement.A series of dilutions of HIV1 RNA were made and used as standards for comparison in each RT-PCR experiment.(https://www.daowen.com)