Exontrap Cloning Vector
A vector with splicing function for the selective cloning of exons
APPLICATIONS
The Exontrap vector has an intrinsic splicing function, allowing selective cloning of exon sequences from large genomic DNA fragments. This new route for the identification of eukaryotic genes does not involve an initial isolation of cellular mRNA. Thus genes that are not transcribed during certain life cycle stages can be identified. Exon/intron mapping is greatly facilitated, since for the determination of exon bounderies only the trapped exons have to be sequenced and compared to the known gene. Also, an exon library can be derived and screened for cell type-specific genes with labeled cDNA from a panel of differentiated cells. The Exontrap function is based on a shuttle vector containing prokaryotic and eukaryotic genetic elements for replication in both bacteria and cell cultures. The vector contains a 5' and 3' exon separated by a 600 bp intron sequence, which contains a polylinker for cloning. The recombinant vector is transfected into eukaryotic cells (e.g. COS cells) and transcribed. The mRNA is processed, i.e. the intron sequences originating from the vector, as well as those being introduced, are removed. After total RNA isolation, the mature mRNA is reverse transcribed into cDNA using a specific primer complementary to a sequence of the bordering exon. The cDNA is amplified by PCR using specific primers, which create restriction sites for further subcloning.
References:
| ORDER INFORMATION | |||
| Cat. No. | Description | Size/Vol. | List |
| 5300-550 | Exontrap kit | 10 µg pET01 | $590.00 |
| Kit includes: cDNA primer 1, 5' PCR primer 2, 3' PCR primer 3 | |||
| Oligos for sequencing: | |||
| 5300-551 | 5' sequencing primer 4 | ||
| 5300-552 | 3' sequencing primer 5 | ||
| Oligos for Exon probes: | |||
| 5300-553 | 5' PCR primer 6 | ||
| 5300-554 | 3' PCR primer 7 | ||
| Shipped at RT; store at 4°C A detailed handbook is provided with the kit | |||