| Popis: |
Publisher Summary Formamide low-temperature polymerase chain reaction (FoLT PCR) is a rapid PCR protocol that allows direct amplification of DNA from whole blood and tissues without any preparative steps—that is, the raw material can be put directly into the tube containing the PCR mixture. It entails the use of 18% formamide, reduced incubation temperatures (80°, 40°, and 60°C), and Thermus thermophilus (Tth) DNA polymerase. This technique has been applied to amplify histocompatibility genes and enzyme genes in several species, including mice and humans. Blood and solid tissues have been used successfully. This chapter demonstrates its use in screening transgenic mice. FoLT PCR saves considerable time and is economical when compared with competing methodologies. Another important characteristic of the FoLT PCR protocol is that even though it is a rapid direct method, it does not compromise sensitivity as compared to protocols that involve DNA purification. One of the advantages of FoLT PCR is that it is suitable for use with most widely used anticoagulants. The only anticoagulants that are not recommended for routine use are lithium heparin, fluoride heparin, and sodium citrate. |
