Minielute purification kit

Manufactured by Qiagen

The MiniElute purification kit is a laboratory equipment product designed for the rapid purification of DNA or RNA samples. It utilizes a silica-based membrane technology to selectively bind nucleic acids, allowing for efficient removal of contaminants and salts. The kit provides a simple and streamlined protocol for DNA or RNA purification, making it a versatile tool for various molecular biology applications.

Automatically generated - may contain errors

Lab products found in correlation

Genemarker v1, by SoftGenetics (1 mentions) Miseq 2 150, by Illumina (1 mentions) Quant it dsdna assay kit, by Thermo Fisher Scientific (1 mentions) Nebnext ultra 2 dna library prep kit, by Illumina (1 mentions) Nextera dna sample preparation kit, by Illumina (1 mentions) Anti flag antibody, by Merck Group (1 mentions) Effectene transfection kit, by Qiagen (1 mentions) Cvcl z232, by Thermo Fisher Scientific (1 mentions) Am9260g, by Thermo Fisher Scientific (1 mentions) Am9759, by Thermo Fisher Scientific (1 mentions) Eo0491, by Thermo Fisher Scientific (1 mentions)

5 protocols using minielute purification kit

DNase I Footprinting of DNA-Protein Interactions

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

DNase I footprinting experiments were performed in TKMC buffer (10 mM Tris, 10 mM KCl, 5 mM MgCl₂ and 5 mM CaCl₂) with 10 mM CHAPS as previously described [26 (link)]. DNA concentrations were adjusted between 200 pM and 1 nM as needed. The reaction was initiated with 0.01-0.1 U DNase I; the amount of DNase I was dependent on the age of the enzyme, the DNA fragment, and DNA concentration. Reaction products were purified using the Qiagen MiniElute purification kit. Capillary electrophoresis (CE) analyses of samples were conducted at the DNA Core Facility at University of Missouri. Data were processed using Genemarker V1.97 software (Softgenetics LLC, State College, PA). DNase I cleavage products were mapped using Sanger sequencing (USB, Affymetrix, Santa Clara, CA). Peaks in the footprint were normalized to a peak not sensitive to PA concentration and plotted as fraction bound vs. PA concentration; the data were fit to a binding isotherm as previously described [26 (link)].

Vasilieva E., Niederschulte J., Song Y., Harris GD J.r., Koeller K.J., Liao P., Bashkin J.K, & Dupureur C.M. (2016). Interactions of Two Large Antiviral Polyamides with the Long Control Region of HPV16. Biochimie, 127, 103-114.

+ Open protocol

+ Expand

Nucleosome Mapping by MNase-seq

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The nucleosome library (0.2 pmol/µl) is digested by MNase (0.05 U/µl) in nuclease digestion buffer (10 mM Tris–HCl, pH 8.0, 2 mM CaCl₂) for a time course of 0 (no MNase used), 5 and 10 min at 37°C. After the defined incubation time, digestion was stopped (2% SDS, 40 mM EDTA). Proteinase K (16 µg) is added to each sample, and the reaction is incubated at 55°C for 1 h. The DNA is purified from the reaction and concentrated using a Qiagen MiniElute Purification Kit. The DNA concentration of each sample is determined by the Invitrogen Quant-iT dsDNA Assay Kit and equalized. Illumina sequencing libraries were generated using NEBNext Ultra II DNA library prep kit. Individual samples are multiplexed and sequenced on an Illumina MiSeq 2 × 150. MNase-seq sequencing results are quality filtered (q > 30) and adapter trimmed using Cutadapt (26 ). The quality reads are merged and mapped to the 7500 nucleosome library sequences using Vsearch (27 (link)). The read counts and end positions are used to determine MNase protection, which is a measurement of the percentage of reads for a specific nucleosome base pair location. MNase protection is calculated for each base pair as the ratio of base pair coverage/total reads for that specific nucleosome.

Handelmann C.R., Tsompana M., Samudrala R, & Buck M.J. (2023). The impact of nucleosome structure on CRISPR/Cas9 fidelity. Nucleic Acids Research, 51(5), 2333-2344.

+ Open protocol

+ Expand

ATAC-seq from Sorted Basal Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For ATAC-seq, 100000 sorted basal cells were collected in 1 ml PBS supplemented with 3% FBS at 4°C. Cells were centrifuged and cell pellets were resuspended in 100 μl lysis buffer (TrisHCl 10 mM, NaCl 10 mM, MgCl2 3 mM, Igepal 0.1%) and centrifuged at 500g for 25min at 4°C. Supernatant was carefully discarded and nuclei were resuspended in 50 μl reaction buffer (Tn5 transposase 2.5 μl, TD buffer 22.5 μl, from Nextera DNA sample preparation kit, Illumina, and 25 μl H₂0). The reaction was performed at 37 °C for 30 min and was stopped by adding 5 μl clean up buffer (NaCl 900 mM, EDTA 300 mM). DNA was purified using the MiniElute purification kit (QIAGEN) following the manufacturer’s protocol. DNA libraries were PCR amplified (Nextera DNA sample preparation kit, Illumina), and size selected from 200 to 800 bp (BluePippin, Sage Sciences), following the manufacturer’s recommendations.

Aragona M., Sifrim A., Malfait M., Song Y., Van Herck J., Dekoninck S., Gargouri S., Lapouge G., Swedlund B., Dubois C., Baatsen P., Vints K., Han S., Tissir F., Voet T., Simons B.D, & Blanpain C. (2020). Mechanisms of stretch-mediated skin expansion at single-cell resolution. Nature, 584(7820), 268-273.

+ Open protocol

+ Expand

ChIP Assay for Flag-jumu Protein Binding

Check if the same lab product or an alternative is used in the 5 most similar protocols

ChIP assays were performed as described previously (Kim et al., 2005 (link)). In brief, S2 cells (CVCL_Z232, Invitrogen, Cat#R690-07) were transfected with the pMK33-Flag-jumu full CDS construct using the Effectene Transfection kit (Qiagen). S2 cells stably expressing Flag-jumu were fixed in 1% formaldehyde at room temperature. The samples were then sonicated to obtain DNA fragments between 200 and 800 bp in length. The fragmented chromatin was incubated with the anti-Flag antibody (Sigma) or IgG (Santa Cruz) bound to Salmon Sperm DNA/Protein A Agarose Slurry (Upstate) overnight at 4°C. The immunoprecipitated DNA fragments were recovered using a MiniElute purification kit (Qiagen) and analyzed by PCR amplification. Similar results were obtained in three independent ChIP experiments. The primer sequences are shown in Supplementary file 1.

Hao Y, & Jin L.H. (2017). Dual role for Jumu in the control of hematopoietic progenitors in the Drosophila lymph gland. eLife, 6, e25094.

+ Open protocol

+ Expand

Genomic DNA Extraction from FFPE and Frozen Nuclei

Check if the same lab product or an alternative is used in the 5 most similar protocols

For FFPE-ATAC samples, single nuclei were isolated following the nuclei isolation protocol stated in the section on nuclei isolation from FFPE tissue sections. For frozen samples, nuclei were isolated following the nuclei isolation protocol in the section on standard ATAC-seq on frozen tissue. For genomic DNA purification, 1 million isolated nuclei were spined down at 3000g for 10 min and then resuspended with 100 µL of lysis buffer (50 mM Tris-HCl at pH 7.5 [Invitrogen 15567027], 1 mM EDTA [Invitrogen AM9260G], 1% SDS [Invitrogen 1553-035], 200 mM NaCl [Invitrogen AM9759], and 200 µg/mL Proteinase K [Thermo Fisher Scientific EO0491]). Nuclei suspension was incubated overnight at 65°C with 1200 rpm shaking in a heat block. On the next day, the mixture was purified with a Qiagen MiniElute purification kit (Qiagen 28004) and eluted in 20 µL of elution buffer. Purified genomic DNA was measured and was run on a 1.5% agarose gel (Lonza 50004) to check size distribution.

Zhang H., Polavarapu V.K., Xing P., Zhao M., Mathot L., Zhao L., Rosen G., Swartling F.J., Sjöblom T, & Chen X. (2022). Profiling chromatin accessibility in formalin-fixed paraffin-embedded samples. Genome Research, 32(1), 150-161.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!