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General description
Optimized one-tube procedure for the synthesis of double-stranded cDNA up to 3 kb from total RNA or mRNA.
The cDNA Synthesis System uses an optimized one-tube procedure, based on the method of Gubler and Hoffmann.
Sample: 1 - 20 μg total RNA, 0.3 - 2 μg mRNA
Assay time: 60 minutes for 1st strand synthesis, 120 minutes for 2nd strand synthesis
Product: double-stranded cDNA copy of an RNA sample
Sample: 1 - 20 μg total RNA, 0.3 - 2 μg mRNA
Assay time: 60 minutes for 1st strand synthesis, 120 minutes for 2nd strand synthesis
Product: double-stranded cDNA copy of an RNA sample
Application
Optimized one-tube procedure for the synthesis of double-stranded cDNA from total RNA or mRNA.
The ds cDNA can be used for the construction of non-directional cDNA libraries, as a starting point for substractive hybridization experiments to enrich differentially expressed genes, and for the in vitro transcription of whole cDNA populations to generate labeled cRNA for hybridization on DNA microarrays.
The ds cDNA can be used for the construction of non-directional cDNA libraries, as a starting point for substractive hybridization experiments to enrich differentially expressed genes, and for the in vitro transcription of whole cDNA populations to generate labeled cRNA for hybridization on DNA microarrays.
Components
- AMV Reverse Transcriptase Buffer, 5x concentrated
- AMV Reverse Transcriptase (25 U/μl)
- 1,4-Dithiothreithol (0.1 M)
- Protector RNase Inhibitor (25 U/μl)
- Oligo (dT)15 primer, 200 μM (1 μg/μl)
- Oligo [(dT24 )T7promotor]65 primer, 100 μM (2 μg/μl)
- dNTP mixture (each nucleotide, 10 mM)
- Control neo mRNA (0.2 μg/μl)
- 2nd strand synthesis buffer, 5x concentrated
- 2nd strand enzyme blend (mixture of DNA Polymerase I, E. coli Ligase and RNase H)
- T4 DNA Polymerase (1 U/μl)
- Double-distilled water
- RNase I (10 U/μl)
- Proteinase K, recombinant, PCR grade (50 U/mll)
Other Notes
For life science research only. Not for use in diagnostic procedures.
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Eye Irrit. 2 - Resp. Sens. 1 - Skin Sens. 1
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
does not flash
Flash Point(C)
does not flash
Certificate of Analysis
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Certificate of Origin
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Product Information Sheet
User Guide
Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.
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