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- W2020886670 abstract "Laser microdissection (LM) provides a useful method for isolating specific cells or tissues from biological samples. Here, we adapted microdissection protocols to allow high-resolution transcript analysis of different tissues from developing Arabidopsis seed. Sufficient RNA (<mml:math xmlns:mml=http://www.w3.org/1998/Math/MathML><mml:mo>∼</mml:mo></mml:math>50 ng) was extracted from endosperm tissue for RT-PCR. However, to obtain enough RNA for microarray analyses, it was necessary to amplify the RNA. PCR- and IVT-based amplification methods were investigated and several important technical aspects of amplification were identified (such as target truncation and alterations in signal intensity). We found that when starting from only 50 ng of RNA, amplification methods based on PCR and IVT produced sufficient product for reliable microarray hybridizations, with two-round IVT giving the best results. Microarray analyses, using endosperm-derived RNA amplified by two-round IVT, reproducibly identified endosperm enriched marker genes. Thus, when combined with RNA-amplification protocols, LM is a robust and reliable technique for high-throughput tissue-specific gene expression analysis." @default.
- W2020886670 created "2016-06-24" @default.
- W2020886670 creator A5039965367 @default.
- W2020886670 creator A5060698730 @default.
- W2020886670 creator A5084224371 @default.
- W2020886670 date "2007-04-05" @default.
- W2020886670 modified "2023-10-15" @default.
- W2020886670 title "Evaluation of Global RNA Amplification and Its Use for High-Throughput Transcript Analysis of Laser-Microdissected Endosperm" @default.
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- W2020886670 doi "https://doi.org/10.1155/2007/61028" @default.
- W2020886670 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/1939914" @default.
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