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For general laboratory use. Not for use indiagnostic procedures. FOR
IN VITRO 
USE ONLY.
 Titan One Tube RT-PCR Kit
Cat. No. 11 939 823 001
50 reactions including 10 control reactionsStore at
15 to
25°C
Store vial 4 (Humancontrol RNA) at
60°Cor below.
Instruction Manual 
Version December 2005
 
Roche Applied Science
2
1.Preface
1.1Table of contents1.Preface .....................................................................................................................................................2
1.1Table of contents...................................................................................................................................................................21.2Kit contents.............................................................................................................................................................................31.3Kit storage/stability...............................................................................................................................................................3
2. Product overview....................................................................................................................................4
2.1Background information.....................................................................................................................................................5
3.Procedures and required materials....................................................................................................6
3.1Before you begin...................................................................................................................................................................63.2Parameters influencing the RT-PCR reaction.............................................................................................................83.3Preparation of Master Mix 1 + 2.....................................................................................................................................93.4RT-PCR Protocol..................................................................................................................................................................10
4. Results ...................................................................................................................................................125.Appendix ................................................................................................................................................15
5.1 Trouble shooting.................................................................................................................................................................155.2References.............................................................................................................................................................................155.3Ordering Information.........................................................................................................................................................15
 
Roche Applied Science
3
1.2Kit content1.3 Kit storage/stability
The kit is stable at -15 to -25°C until the control date printed on the label.
Note: 
The supplied control RNA should be stored separately at -60°C or below.
Bottle/CapLabelContentincluding function
1Titan enzyme mix•50 µl•Titan enzyme mix (AMV and Expand HighFidelity) in storage buffer •1 reaction/µl2dNTP mix•200 µl•10 mM total (2.5 mM each dNTP)3RNase Inhibitor•50 µl•[5 U/µl]4Human control RNA•50 µl•[2 pg/µl]•K562 total RNA with MS2 carrier RNA5Control primer mix•20 µl•20 µM (total)•human ß-actin upstream and downstream primer 6H
2
0 redist., PCR grade2 x 1 mlRT-PCR reactionbuffer •1 ml 5x conc.•with 7.5 mM MgCl
2
and DMSODTT solution•1 ml•100 mMMgCl
2
stock solution•1 ml•25 mM
of 00

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