Qpcr

Fast SYBR® Green Cells-to-CT™ Kit
(Part Number 4402956, 4402957)

Protocol I. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 A. Product Description and Background B. Materials Provided with the Kit, Storage Conditions, and Safety Information C. Materials Not Provided with the Kit D. Related Products Available from Applied Biosystems Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 A. Suggested Controls and Replicates B. Cell Lysis C. Reverse Transcription (RT) D. Fast Real-Time PCR Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
A. No PCR Product or Unexpected PCR Products B. RT-PCR Products in the Negative Control Reactions

II.

III.

IV.

Appendix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 A. PCR Primer Design Suggestions B. Pilot Experiment C. Purification of RNA from Cells-to-CT Lysates D. Quality Control E. Safety Information E.I. General chemical safety E.II. Safety alerts

P/N 4403786 Revision A Revision Date: September 3, 2008 For research use only. Not for use in diagnostic procedures. By use of this product, you accept the terms and conditions of all applicable Limited Label Licenses. For statement(s) and/or disclaimer(s) applicable to this product, see below. Information in this document is subject to change without notice. Applied Biosystems assumes no responsibility for any errors that may appear in this document. Applied Biosystems disclaims all warranties with respect to this document, expressed or implied, including but not limited to those of merchantability or fitness for a particular purpose. In no event shall Applied Biosystems be liable, whether in contract, tort, warranty, or under any statute or on any other basis for special,...