The Nucleic Acid Mini Screen is an efficient screen formulated to assist in the determination of preliminary crystallization conditions of nucleic acid fragments.1 The formulation is based upon the publication, “A Highly Efficient 24 Condition Matrix for the Crystallization of Nucleic Acid Fragments” where the preliminary crystallization conditions of 35 nucleic acids were determined utilizing this formulation.1
Samples include DNA-Drug complexes, C-Tetrad and G-Quartet Motifs, RNA oligomers, and others. By using 1 to 4 mM oligonucleotide stock concentration, the screen requires less than 100 µl of sample. The screen is typically performed at 4°C although room temperature incubations can also be performed. To evaluate the effect of equilibration kinetics as well as initial and final sample concentrations, the screens are typically performed using the hanging or sitting drop vapor diffusion method with two drops on a slide (1 µl + 2 µl and 2 µl + 2 µl), side by side. The 24 well format of the mini screen provides for a fast setup and uses small amounts of sample. This makes it possible to cost-effectively screen many sequences as well as variations of a particular sequence. The composition of the Nucleic Acid Mini Screen allows one to apply the formulation to other nucleic acids such as deoxy- and ribozymes, pseudoknots, and tRNAs.
Each Nucleic Acid Mini Screen kit (HR2-118) contains 24 unique reagents, 1.0 ml each. All solutions are formulated using ultra-pure Type 1 water and are sterile filtered. Dehydrants is available separately as 200 ml of 35% v/v (+/-)-2-Methyl-2,4-pentanediol in deionized water(catalog number HR2-863). Formulated in Type 1+ ultrapure water: 18.2 megaohm-cm resistivity at 25°C, < 5 ppb Total Organic Carbon, bacteria free (<1 Bacteria (CFU/ml)), pyrogen free (<0.03 Endotoxin (EU/ml)), RNase-free (< 0.01 ng/mL) and DNase-free (< 4 pg/µL) and sterile filtered into sterile containers.
Note: Crystallization kit (HR2-118) and Dehydrant/reservoir solution (HR2-863) are sold separately. Both items are needed to perform the screen.
1. Berger, et al., A Highly Effective 24 Condition Matrix for the Crystallization of Nucleic Acid Fragments. Acta Cryst. Section D. (1996) Vol. D52 Part 3, 465-468.
2. Adams, A., Nucleic Acids Research (2002) Vol. 30, 719-725.
3. Crystallization and preliminary X-ray diffraction analysis of an Escherichia coli tRNAGly acceptor-stem microhelix. C. Forster, M. Perbandt, A. B. E. Brauer, S. Brode, J. P. Furste, C. Betzel and V. A. Erdmann. Acta Cryst. (2007). F63, 46-48.