Description
- Reagents to synthesize double-stranded cDNA from up to 40µg of mRNA
- Oligo(dT)15 Primer and Random Primers included for choice of priming methods
- Materials provided for 10 adaptor ligation reactions
The Universal RiboClone™ cDNA Synthesis System is used for double-stranded cDNA synthesis from RNA. The system also includes materials for ligation of the cDNA into a suitable vector. First-strand synthesis is driven by Avian Myeloblastosis Virus (AMV) Reverse Transcriptase and either Random Primers or Oligo(dT)15 Primer. This is followed directly by second-strand synthesis using RNase H and DNA Polymerase I. After treatment with T4 DNA Polymerase to create blunt ends, the double-stranded cDNA molecules are prepared for cloning by size fractionation and addition of EcoRI Adaptors. The resulting cDNA preparation is ready for cloning into a suitable vector.
Protocols
What’s in the box?
| Item/ Catalog number selected: C4360 | Part # | Size | Concentration | Available Separately |
|---|---|---|---|---|
| Sodium Pyrophosphate | C113A | 1 × 50μl | 40mM | |
| First Strand 5X Buffer | C121A | 1 × 200μl | ||
| T4 DNA Ligase 10X Buffer | C126A | 1 × 500μl | View Product | |
| Spin Columns | C1281 | 1 × 10 each | View Product | |
| EcoRI Adaptors | C129A | 1 × 150pmol | View Product | |
| Kinase 10X Buffer | C131A | 1 × 500μl | View Product | |
| ATP | C134A | 1 × 50μl | ||
| 1.2kb Kanamycin Positive Control RNA | C138B | 1 × 10μg | 0.5mg/ml | View Product |
| Oligo(dT)15 Primer | C144A | 1 × 20μg | 500μg/ml | View Product |
| Random Primers | C146A | 1 × 20μg | View Product | |
| Second Strand 2.5X Buffer, AMV | C630A | 1 × 800μl | ||
| T4 DNA Ligase | M180A | 1 × 100u | ||
| DNA Polymerase I | M205A | 1 × 500u | ||
| T4 Polynucleotide Kinase | M410A | 1 × 100u | View Product | |
| T4 DNA Polymerase | M421A | 1 × 100u | ||
| RNase H | M428B | 1 × 20u | ||
| AMV Reverse Transcriptase (HC) | M900A | 1 × 600u | View Product | |
| Recombinant RNasin® Ribonuclease Inhibitor | N251E | 1 × 1,000u | ||
| Nuclease-Free Water | P119A | 1 × 1,250μl | View Product | |
| Acetylated BSA | R946A | 1 × 400μl | View Product | |
| Sephacryl® S-400 | V318A | 1 × 10ml | View Product |
SDS For C4360
Download SDSUse Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.
Storage Conditions
See Protocol for detailed storage recommendations.
