The Rapid-Screen™ Arrayed cDNA Library system utilizes two
separate plates - a "Master Plate" and a "Sub- Plate"
- to isolate the clone of interest. The Master Plate is a standard
96-well microtiter plate arrayed with a size-selected cDNA library
at a titer of approximately 5000 clones per well. The Master Plate
is screened using gene-specific primers followed by agarose electrophoresis
of the PCR products.
Depending on the abundance of the gene of interest, positive wells
are subsequently identified via corresponding bands on agarose gel.
For each well of the Master Plate there is a corresponding Sub-Plate.
Each Sub-Plate is arrayed at a titer of 50 clones per well. Gene-specific
PCR is performed on the Sub-Plate to identify positive wells. Sub-plate
clones are housed in E.coli. A small aliquot from the positive well(s)
of the Sub-Plate are plated onto standard LB agar and grown overnight.
Using colony PCR, 96-colonies are screened with an average of approximately
two positive clones per positive Sub-Plate well. These clones are
easily amplified for further analysis.
- 96-well arrayed cDNA library Master Plate (5000 clones/well)
- Rapid-Screen vector primer pCMV6-XL4 (10 pmol/?l)
- Rapid-Screen positive control (483 bp)
- Quanti-Ladder™ DNA Marker - a ready-to-use molecular
marker for easy quantitation and fragment size determination (100
?l for 20 lanes)
- Four 96-well plate re-sealing tapes
- 96-well arrayed cDNA library Sub-Plate (50 clones/well).