Clontech PCR-SelectTM Differential Screening Kit

Description

Complete kit for differential screening of subtracted libraries obtained using the Clontech PCR-Select cDNA Subtraction Kit. The differential screening procedure can be used to identify putative differentially expressed sequences in the subtracted library before performing Northern blot analysis. The kit contains reagents for making cDNA probes for differential screening and hybridization control cDNAs. 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 The PCR-Select Differential Screening Kit detects rare, differentially expressed cDNAs.The SMART PCR cDNA Synthesis Kit Cat. # 634902 was used to preamplify total RNA from a gamma-globin-producing cell line and a beta-globin-producing cell line. PCR-Select cDNA subtraction was performed using the gamma-line cDNA as tester and beta-line cDNA as driver; for the reverse subtraction, tester and driver were switched. The subtracted cDNA was then cloned, and randomly selected clones spotted on nylon membranes for duplicate screening. Membranes were hybridized with the indicated probe. 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