Pools and Superpools
Amplicon Express' patent pending Pool & Superpool technology is used to screen your library for clones of interest, much like our
Nylon Filter sets, though the significant advantage of our Pools and Superpools sets are that they can be used to
screen your libraries up to 3000 times versus 3-5 times for nylon filters, and they represent one of the most powerful and accurate ways for you
to find your region of interest within that library. Amplicon Express has also developed proprietary software to automate analysis of your Pool
and Superpool results and help to eliminate error in interpreting results.
Amplicon Express' Pooling & Superpooling System
This pooling and superpooling system enables researchers to use PCR for screening a BAC library and identify the specific plate & well
containing a BAC clone with a sequence of interest. The screening is done in two separate rounds of PCR on pooled BAC clones (Round I
and Round II). Round I identifies which superpool(s) have hits with the PCR signal and then the positive superpools are followed up in
Round II to identify the plate and well position of the positive BAC clone(s).
Each kit is custom built for the researcher and the total number of Superpools in the kit will depend on the total number of BAC clones in the library. Each Superpool will have a corresponding 96-well plate containing all corresponding Plate Pools, Row Pools, Column Pools that have been matrixed together to reduce the number of PCR experiments (called a Matrix Output Plate). The entire resource is aliquoted into TWO identical working stock sets, to help reduce the risk of contaminating the entire resource or for multiple researchers to utilize the resource.
The Round I PCR is performed on all the superpools (containing all BAC clones in the library). Each Superpool contains 2,688 individual BAC clones from 7 plates. The results from Round I PCR will identify which Superpool(s) contains BAC clone(s) with the sequence of interest (there may be more than one Superpool identified). The researcher may pursue one or more positive hits found in Round I PCR.
The Round II PCR is performed on the Matrixed Plate, Row, Column pools for the specific Superpool being investigated. The Round II requires only 23 PCR experiments plus controls. The results from Round II of PCR should allow the researcher to identify the exact plate and well position for the positive hit in the particular Superpool under investigation. This allows the researcher to have identified the specific BAC clone or clones of interest, out of the complete BAC library, that have the sequence where the PCR primers were targeted at.
Each kit is custom built for the researcher and the total number of Superpools in the kit will depend on the total number of BAC clones in the library. Each Superpool will have a corresponding 96-well plate containing all corresponding Plate Pools, Row Pools, Column Pools that have been matrixed together to reduce the number of PCR experiments (called a Matrix Output Plate). The entire resource is aliquoted into TWO identical working stock sets, to help reduce the risk of contaminating the entire resource or for multiple researchers to utilize the resource.
The Round I PCR is performed on all the superpools (containing all BAC clones in the library). Each Superpool contains 2,688 individual BAC clones from 7 plates. The results from Round I PCR will identify which Superpool(s) contains BAC clone(s) with the sequence of interest (there may be more than one Superpool identified). The researcher may pursue one or more positive hits found in Round I PCR.
The Round II PCR is performed on the Matrixed Plate, Row, Column pools for the specific Superpool being investigated. The Round II requires only 23 PCR experiments plus controls. The results from Round II of PCR should allow the researcher to identify the exact plate and well position for the positive hit in the particular Superpool under investigation. This allows the researcher to have identified the specific BAC clone or clones of interest, out of the complete BAC library, that have the sequence where the PCR primers were targeted at.