Nucleic Acid Isolation.indd - Invitrogen

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A Wide Variety of Applications Fig. 22. Isolation of tRNA-VAL from rat mitochondria. S: supernatant after capture, W: washing solution, E: eluate, position of the isolated full-length tRNA-VAL is indicated by the arrow. Courtesy of M. Mørl, University of Munich, Germany. Ordering Information Specific Capture of Nucleic Acids Double-stranded DNA fragments immobilised to Dynabeads ® Streptavidin are easily converted to single-stranded DNA templates or probes (fig. 19). The bead-bound sequences can then be used for solid-phase hybridisation capture of specific DNA/RNA sequences (fig. 22). Streptavidin-coupled Dynabeads ® provide an efficient solid-phase alternative to nitrocellulose. An inherent benefit of magnetic handling is that downstream manipulations and buffer changes can be done by simply concentrating the bead-bound target at the tube-wall by using a magnet. The excellent reaction kinetics of the Dynabeads ® is comparable to liquid phase kinetics. A singlestranded biotinylated probe can be bound directly to the beads for capture of target sequences. Alternatively, an indirect capture approach can be used, as the reaction kinetics are faster, in some cases, when the biotinylated probe is added to the sample for target hybridisation, prior to immobilisation to the beads. The 1.0 µm Dynabeads ® MyOne Streptavidin C1 have a very high surface area per mg of beads, enabling high enrichment of low abundance DNA/RNA. The slightly negatively charged surfaces of both the Dynabeads ® M-270 Streptavidin and the Dynabeads ® MyOne Streptavidin C1 also ensure negligible non-specific binding of non-target DNA sequences. Examples of applications include; sequence-specific capture, cDNA selection and enrichment, isolation of cell-specific transcripts and mRNA differential display. A reference list is available upon request. Product Volume Product No. Dynabeads ® MyOne Streptavidin C1 Dynabeads ® M-270 Streptavidin Dynabeads ® M-280 Streptavidin Dynabeads ® kilobaseBINDER Dynabeads ® M-280 Streptavidin, Binding Solution and Washing Solution. Please see chapter 7 for additional Dynabeads ® Streptavidin products. 84 2 ml (10 mg/ml) 10 ml (10 mg/ml) 100 ml (10 mg/ml) 2 ml (10 mg/ml) 10 ml (10 mg/ml) 100 ml (10 mg/ml) 2 ml (10 mg/ml) 10 ml (10 mg/ml) 100 ml (10 mg/ml) 650.01 650.02 650.03 653.05 653.06 653.07 112.05 112.06 602.10 200 isolations 601.01 www.dynalbiotech.com
Dye Terminator Removal Removal of unincorporated dye terminators, salts and other unwanted components from the extension products is critical for both standard and capillary DNA sequencing systems. Dynabeads ® Sequencing Clean-Up is a simple three-step magnetic bead based system for purifying DNA sequencing reactions (fig. 23). The unique uniformity of the beads guarantees consistently high quality, reproducible results, long reading lengths and high Phred20 scores (fig. 24 and 25). Dynabeads ® Sequencing Clean-Up removes dye terminators from 10-20 µl sequencing reactions and is compatible with any sequencing chemistries including including BigDye, dRhodamine dye, Rhodamine, DYEnamic ET and WellRED terminators. After purification, the extension products can be separated on any DNA sequencing instrumentation. Main Advantages of Dynabeads ® Sequencing Clean-Up: • Efficient removal of excess dye terminators. • Fast 15 minutes procedure. • No centrifugation or filtration. • Automated protocols are available (see www.dynalbiotech.com). Ordering Information www.dynalbiotech.com 85 Sequencing Clean-up Fig. 23. Principle for purification of DNA sequencing preparation using Dynabeads ® Sequencing Clean-up. Fig. 24. BAC-clone 1105B13 directly sequenced using BigDye ® Terminator Cycle Sequencing Ready Reaction Kit (ABI). The sequencing products were purified using the sample injected into an ABI PRISM ® 310 automated capillary DNA sequencer. Fig. 25. M13 mp18 + template sequenced in a reaction using DYEnamic TM ET, purified with Dynabeads ® Sequencing Clean-Up and analysed on the MegaBase TM 1000. Product Volume Product No. Dynabeads ® Sequencing Clean-Up 500 samples 661.01 Dynabeads ® Sequencing Clean-Up 2500 samples 661.02 6 7 Nucleic Acid Isolation
Page 1 and 2: 6. Nucleic Acid Isolation Isolation
Page 3 and 4: Nucleic Acid Isolation Isolation of
Page 5 and 6: Direct mRNA isolation with the Dyna
Page 7 and 8: mRNA for cDNA Libraries Dynabeads
Page 9 and 10: Ordering Information www.dynalbiote
Page 11 and 12: A Variety of Sample Types and Downs
Page 13 and 14: Dynabeads ® Streptavidin for Super
Page 15: Isolating RNA and DNA Binding Prote

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