RL-SAGE
Robust LongSage (RL-SAGE) is a version of Serial Analysis of Gene Expression SAGE. It is an improvement over LongSAGE analysis that is less advanced than the latest innovation in SAGE technology, SuperSAGE.
Improvements over previous model
RL-SAGE was developed to solve several of the problems inherent in SAGE and LongSAGE protocols, including small insert sizes and low cloning efficiency. Improvements include the ability to generate a library with an insert size of 50 ng mRNA, much smaller than previous LongSAGE insert size of 2 μg mRNA [1]and using a lower number of ditag polymerase chain reactions (PCR) to obtain a complete cDNA library. [2]
Notable experiments
3,339 novel genes in magnaporthe Grisea were identified using RL-SAGE techniques. [3]
References
- ^ Saha, S., et al. (2002). "Using the transcriptome to annotate the genome." Nat Biotechnol 20(5): 508-512.
- ^ Gowda, M., et al. (2004). "Robust-LongSAGE (RL-SAGE): a substantially improved LongSAGE method for gene discovery and transcriptome analysis." Plant Physiol 134(3): 890-897.
- ^ Gowda, M., et al. (2006). "Deep and comparative analysis of the mycelium and appressorium transcriptomes of Magnaporthe grisea using MPSS, RL-SAGE, and oligoarray methods." BMC Genomics 7: 310.