Clontech PCR-SelectTM cDNA Subtraction Kit

Description

Kit for identifying cDNAs that correspond to differentially expressed sequences in one cDNA population compared with another. Enough reagents are provided for seven cDNA reactions. PCR primers are provided for 50 primary and 100 secondary PCR amplificationsenough for one control and six complete subtractions if the cDNA from each synthesis is used for tester and driver in separate subtractions. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back Southern blot analysis showing enrichment of a gene activated in human Jurkat T-cells by PHA/PMA treatment, and reduction of an abundant housekeeping gene in subtracted cDNA.Tester cDNA was prepared from human Jurkat leukemic T-cells and incubated with 2 mug/ml HA and 2 ng/ml PMA for 72 hr. Driver cDNA was prepared from the same untreated cells. Amplified tester, driver, and subtracted cDNA were electrophoresed on a 1.5 % agarose gel 0.3 mug per lane , transferred onto nylon filters, and hybridized with either an IL-2R alpha probe, a known marker of activation Panel A , or a G3PDH housekeeping gene probe Panel B . Back Required Products This kit is based on Takaras Advantage 2 Polymerase Mix. It contains all reagents needed for a hot-start PCR, including control template and primer mix. This kit is ideal for PCR applications that require high fidelity, such as cDNA amplification or library construction. Enough reagents are supplied for 100 PCR reactions of 50 mul each. Back Back Back Advantage 2 Polymerase Mix is a hot-start PCR polymerase mix that contains Takaras TitaniumTaqDNA Polymerase with TaqStart Antibody , and a minor amount of a proofreading polymerase for a balance of higher yield and higher fidelity than wild-typeTaq. The Mix is ideal for amplification of long templates up to 18 kb and complex genomic DNA up to 6 kb. Two different optimized buffers are supplied with the polymerase mix. dNTPs need to be purchased separately. Enough enzyme mix and PCR buffer are supplied for 100 PCR reactions of 50 mul each. Back Back Back A monoclonal antibody isotype IgG2b that specifically binds to and inactivatesTaqDNA polymerase below 70C for use in hot-start PCR. PCR with TaqStart Antibody has been proven to prevent generation of nonspecific amplification products and primer-dimer artifacts. Back Back Back