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Enzymatic amplification of DNA by PCR: standard procedures and optimization.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Kramer, Martha F. Coen, Donald M. |
| Copyright Year | 2006 |
| Abstract | This unit describes a method for amplifying DNA enzymatically by the polymerase chain reaction (PCR), including procedures to quickly determine conditions for successful amplification of the sequence and primer sets of interest, and to optimize for specificity, sensitivity, and yield. The first step of PCR simply entails mixing template DNA, two appropriate oligonucleotide primers, Taq or other thermostable DNA polymerases, deoxyribonucleoside triphosphates (dNTPs), and a buffer. Once assembled, the mixture is cycled many times (usually 30) through temperatures that permit denaturation, annealing, and synthesis to exponentially amplify a product of specific size and sequence. The PCR products are then displayed on an appropriate gel and examined for yield and specificity. Recommended optimization conditions are included. |
| Starting Page | 4 |
| Ending Page | 9 |
| Page Count | 6 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | http://media.wiley.com/CurrentProtocols/047150338X/047150338X-sampleUnit.pdf |
| PubMed reference number | 18770830v1 |
| Alternate Webpage(s) | https://doi.org/10.1002/0471142956.cya03ks37 |
| DOI | 10.1002/0471142956.cya03ks37 |
| Journal | Current protocols in cytometry |
| Volume Number | 3 |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Buffers Clinical Use Template DNA-Directed DNA Polymerase Deoxyribonucleosides Gene Amplification Technique Oligonucleotide Primers Polymerase Chain Reaction purine deoxyribonucleoside triphosphate biosynthetic process |
| Content Type | Text |
| Resource Type | Article |
