( phenol-based method with an added gl...)Hands-free Tissue Disruption No Homogenization Needed

, phenol-based method, with an added glass filter cleanup step) or Kit 2 (GITC, glass fiber filter-based). RNA was isolated following the protocol for each method with the addition of a glass fiber filter purification step for Kit 1. (A) Agilent 2100 bioanalyzer scans demonstrate the high quality total RNA isolated using the MELT Total RNA Isolation System. RIN = RNA Integrity Number, scaled from 110. (B) The averaged yield from 4 replicates with an input of ~7 mg of frozen or fresh mouse tissue.

High Quality RNA=Superior Results

The goal of any RNA isolation procedure is to recover an RNA population that faithfully mirrors the biology of the sample at the time of procurement. Ambion scientists have extensively validated the MELT technology to ensure that the purified RNA is functionally equivalent to RNA isolated using other popular methods. RNA purified using MELT technology, or two competing kits was first evaluated by real-time RT-PCR. Six distinct mRNAs were quantified across five different mouse tissues that were either freshly collected or flash-frozen. No significant difference was observed between samples isolated with MELT technology, or with competitor Kits 1 and 2 (data for 3 of the targets are shown in Figure 3). In addition, normalized microarray signal intensities obtained from samples isolated using MELT revealed higher Percent Present Calls (Figure 4C) and an excellent correlation (Figure 4A and B) compared to samples generated using the method recommended in Affymetrix's GeneChip Expression Analysis Technical Manual (Kit 1 with an added cleanup step).

Figure 3. qRT-PC
'"/>

Source:

Page: All 1 2 3 4 5 6 7
Related biology technology :

1. Innovative Tissue Array Technology for High-Throughput Screening of Gene Expression
2. Detecting Gastrin-Releasing Peptide Receptor by in situ PCR on Archived Tissue
3. Simple Isolation of RNA from Tissue and Cultured Cells
4. Tissue Specificity for Mutation Parallels Tissue Specificity for Cancer
5. Recover High Yields of Total Nucleic Acid from Formalin-fixed, Paraffin-embedded (FFPE) Tissue
6. RNase Activity in Mouse Tissue: Classification, Hierarchy, and Methods for Control
7. Stabilizing RNA in Fresh and Frozen Tissues for RNA Isolation
8. Genomic DNA Preparation from RNAlater Preserved Tissues
9. DNA Isolation From 300 to 450 mg Dried or 600 to 1,200 mg Fresh Leaf Tissue
10. DNA Isolation From 50 to 20 mg of Paraffin-Embedded Tissue
11. Genomic DNA Isolation From 510 mg Marine Invertebrate Tissue