Illustra nap columns

Manufactured by GE Healthcare

Sourced in United States

The Illustra NAP Columns are lab equipment designed for the purification of nucleic acids. They provide a simple and efficient method for the extraction and purification of DNA and RNA from various sample types.

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Lab products found in correlation

Wheat germ agglutinin (wga), by Vector Laboratories (2 mentions) Riboprobe in vitro transcription system, by Promega (1 mentions) Microtof q 2, by Bruker (1 mentions) Wheat germ agglutinin (wga), by GE Healthcare (1 mentions)

Close spelling variants of this product

Illustra™ NAP-25 columns Illustra NAP-5 desalting columns NAP column

4 protocols using illustra nap columns

Biotinylation of Synthesized tRNA

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The pGEM-T Easy plasmids containing the tRNA^Leu_CAG sequences were used as templates and amplified by PCR for the in vitro transcription of tRNA sequences using corresponding primer pairs (Table S2) and Pfu polymerase. After gel extraction, in vitro transcription was performed using the Riboprobe® in vitro Transcription Systems (Promega). The residual DNA was digested using DNase I, and the tRNAs were purified by phenol/chloroform precipitation. After rehydration with RNase-free destilled water (DW), tRNA was eluted using Illustra NAP columns (GE Healthcare, USA).
For the in vitro biotinylation of the synthesized tRNA, we generated eFx flexizyme, according to the protocol described previously,⁴⁰ and thus charged the synthesized tRNA with biotin-Phe-DBE.

Kwon N.H., Lee M.R., Kong J., Park S.K., Hwang B.J., Kim B.G., Lee E.S., Moon H.G, & Kim S. (2018). Transfer-RNA-mediated enhancement of ribosomal proteins S6 kinases signaling for cell proliferation. RNA Biology, 15(4-5), 635-648.

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Oligonucleotide Synthesis and Purification

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All oligonucleotides were synthesized on an ABI 392 DNA/RNA synthesizer at a 1 μmol scale using UltraMild phosphoramidites in combination with UltraMild capping reagents (emp Biotech). 0.3 M BTT (emp Biotech) was used as an activator. Standard coupling protocols with DMTon strategy were used and coupling times for the 5’‐cap phosphoramidites were extended to 12 min. Cleavage from the solid support was carried out with 0.05 M K₂CO₃ solution in MeOH within 4 h. The cleavage solution was then desalted with illustra NAP columns (GE Healthcare) and RNase‐free water. Purification of the resulting oligonucleotide solution was carried out by RP‐HPLC after vacuum concentration. ESI‐MS was performed on a Bruker micrOTOF‐QII.

Kaufmann J., Müller P., Andreadou E, & Heckel A. (2022). Green‐Light Activatable BODIPY and Coumarin 5’‐Caps for Oligonucleotide Photocaging. Chemistry (Weinheim an Der Bergstrasse, Germany), 28(36), e202200477.

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WGA Antibody Labeling Protocol

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2 mg ml⁻¹ WGA (Vector Labs) was incubated for 1 h at RT while shaking with Atto565-NHS esther (Atto-tec) or AbberiorFlip565-NHS esther (Abberior) at a ratio of 1:6 with the pH raised to 8.3 using sodium bicarbonate. The mixture was purified using illustra NAP Columns (GE Healthcare) according to manufacturer′s instructions and eluted with slightly acidic PBS to recover labeled antibody at neutral pH. The protein concentration was estimated by absorption spectrometry to 0.5 mg ml⁻¹ WGA-Atto565 and to ~1 mg ml⁻¹ WGA-AbberiorFlip565.

Grußmayer K.S., Geissbuehler S., Descloux A., Lukes T., Leutenegger M., Radenovic A, & Lasser T. (2020). Spectral cross-cumulants for multicolor super-resolved SOFI imaging. Nature Communications, 11, 3023.

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Fluorescent Labeling of Antibodies and Lectins

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2 mg ml -1 donkey anti-mouse (H+L) highly cross-adsorbed antibody (Invitrogen) and 2 mg ml -1 wheat germ agglutinin (WGA, Vector Labs) was incubated with Abberior FLIP 565-NHS (Abberior) at a ratio of 1: 6 for 1h at RT while shaking with the pH raised to 8.3 using sodium bicarbonate. The mixture was purified using illustra NAP Columns (GE Healthcare) according to manufacturer's instructions and eluted with slightly acidic PBS to recover the labeled antibody at neutral pH. The protein concentration was estimated by absorption spectrometry to <1.5 mg ml -1 for donkey anti-mouse Abberior FLIP 565 and <2.5 mg ml -1 for WGA-Abberior FLIP 565. Widefield (average of the image sequence), 2 n , 3 rd , 4 th , 5 th & 6 th order SOFI processed using the 2D code that extends to higher orders. 50% glycerol in PBS with <0.68 kW cm -2 561nm excitation, 6000 frames at 50 ms exposure time. Scale bar 5 μm.

Grußmayer K., Lukes T., Lasser T, & Radenovic A. (2020). Self-Blinking Dyes Unlock High-Order and Multiplane Super-Resolution Optical Fluctuation Imaging. ACS nano, 14(7).

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