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10.1186/1471-2199-9-77 http://scihub22266oqcxt.onion/10.1186/1471-2199-9-77 18771595!2535778!18771595     free     free     free Deprecated: Implicit conversion from float 235.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 235.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 235.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 235.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 235.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 235.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 235.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       BMC+Mol+Biol 2008 ; 9 (ä): 77 Nephropedia Template TP {{tp|p=18771595|t=2008. Phi29 polymerase based random amplification of viral RNA as an alternative to random RT-PCR |pdf=|usr=}}{{18771595}} gab.com Text Phi29 polymerase based random amplification of viral RNA as an alternative to random RT-PCR JO: BMC Mol Biol http://www.kidney.de/pget.php?&tw=1&pmid=18771595 #FOAvip BACKGROUND: Phi29 polymerase based amplification methods provides amplified DNA with minimal changes in sequence and relative abundance for many biomedical applications. RNA virus detection using microarrays, however, can present a challenge because phi29 DNA polymerase cannot amplify RNA nor small cDNA fragments (<2000 bases) obtained by reverse transcription of certain viral RNA genomes. Therefore, ligation of cDNA fragments is necessary prior phi29 polymerase based amplification. We adapted the QuantiTect Whole Transcriptome Kit (Qiagen) to our purposes and designated the method as Whole Transcriptome Amplification (WTA). RESULTS: WTA successfully amplified cDNA from a panel of RNA viruses representing the diversity of ribovirus genome sizes. We amplified a range of genome copy numbers from 15 to 4 x 10(7) using WTA, which yielded quantities of amplified DNA as high as 1.2 microg/microl or 10(10) target copies. The amplification factor varied between 10(9) and 10(6). We also demonstrated that co-amplification occurred when viral RNA was mixed with bacterial DNA. CONCLUSION: This is the first report in the scientific literature showing that a modified WGA (WTA) approach can be successfully applied to viral genomic RNA of all sizes. Amplifying viral RNA by WTA provides considerably better sensitivity and accuracy of detection compared to random RT-PCR. Twit Text FOAVip Phi29 polymerase based random amplification of viral RNA as an alternative to random RT-PCR ????BMC Mol Biol http://www.kidney.de/pget.php?&tw=1&pmid=18771595 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=18771595 #FOAvip English Wikipedia <ref>{{Cite pmid|18771595}}</ref> Phi29 polymerase based random amplification of viral RNA as an alternative to random RT-PCR #MMPMID18771595 Berthet N; Reinhardt AK; Leclercq I; van Ooyen S; Batejat C; Dickinson P; Stamboliyska R; Old IG; Kong KA; Dacheux L; Bourhy H; Kennedy GC; Korfhage C; Cole ST; Manuguerra JC BMC Mol Biol 2008[Sep]; 9 (ä): 77 PMID18771595show ga BACKGROUND: Phi29 polymerase based amplification methods provides amplified DNA with minimal changes in sequence and relative abundance for many biomedical applications. RNA virus detection using microarrays, however, can present a challenge because phi29 DNA polymerase cannot amplify RNA nor small cDNA fragments (<2000 bases) obtained by reverse transcription of certain viral RNA genomes. Therefore, ligation of cDNA fragments is necessary prior phi29 polymerase based amplification. We adapted the QuantiTect Whole Transcriptome Kit (Qiagen) to our purposes and designated the method as Whole Transcriptome Amplification (WTA). RESULTS: WTA successfully amplified cDNA from a panel of RNA viruses representing the diversity of ribovirus genome sizes. We amplified a range of genome copy numbers from 15 to 4 x 10(7) using WTA, which yielded quantities of amplified DNA as high as 1.2 microg/microl or 10(10) target copies. The amplification factor varied between 10(9) and 10(6). We also demonstrated that co-amplification occurred when viral RNA was mixed with bacterial DNA. CONCLUSION: This is the first report in the scientific literature showing that a modified WGA (WTA) approach can be successfully applied to viral genomic RNA of all sizes. Amplifying viral RNA by WTA provides considerably better sensitivity and accuracy of detection compared to random RT-PCR. |*DNA-Directed DNA Polymerase[MESH] |Bacillus Phages/*enzymology[MESH] |Base Sequence[MESH] |DNA Primers/genetics[MESH] |DNA, Bacterial/genetics[MESH] |Gene Expression Profiling/*methods[MESH] |Genome, Viral[MESH] |Genomics[MESH] |Nucleic Acid Amplification Techniques/*methods[MESH] |Oligonucleotide Array Sequence Analysis[MESH] |RNA Viruses/genetics[MESH] |RNA, Viral/*genetics[MESH] |Reverse Transcriptase Polymerase Chain Reaction/methods[MESH] |Rift Valley fever virus/genetics[MESH]Phi29 polymerase based random amplification of viral RNA as an alterna(epmc).pdf DeepDyve Pubget Overpricing