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Amplification of cDNA generated by reverse transcription of mRNA.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Sambrook, Joseph Russell, David Williams |
| Copyright Year | 2006 |
| Abstract | INTRODUCTION In this method, an oligodeoxynucleotide primer hybridized to mRNA is extended by an RNA-dependent DNA polymerase to create a cDNA copy that can be amplified by PCR. Depending on the purpose of the experiment, the primer for first-strand cDNA synthesis can be specifically designed to hybridize to a particular target gene, or a general primer such as oligo(dT) can be used to prime cDNA synthesis from essentially all mammalian mRNAs. Similarly, the reverse primer used in the subsequent amplification reaction can be gene-specific or general (e.g., random hexamers). To reduce the chance of contamination with exogenous DNAs, prepare and use a special set of reagents and solutions for PCR only. Bake all glassware for 6 hours at 150°C and autoclave all plasticware. |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | http://vetbiotech.um.ac.ir/parameters/vetbiotech/filemanager/new_admin/Protocols/PCR/Amplification%20of%20cDNA%20Generated%20by%20Reverse%20Transcription%20of%20.pdf |
| PubMed reference number | 22485327v1 |
| Alternate Webpage(s) | https://doi.org/10.1101/pdb.prot3837 |
| DOI | 10.1101/pdb.prot3837 |
| Journal | CSH protocols |
| Volume Number | 2006 |
| Issue Number | 1 |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Autoclave DNA, Complementary Gene Amplification Technique Mammals Oligonucleotide Primers RNA Reagents Reverse Transcription Solutions |
| Content Type | Text |
| Resource Type | Article |
