Log In Sign up
The largest database of trusted experimental protocols

Dig luminescent detection kit

Manufactured by Merck Group
Visit Supplier
Sourced in United States

The DIG luminescent detection kit is a laboratory equipment product used to detect and quantify the presence of digoxigenin (DIG) labeled molecules. The kit provides the necessary reagents and components to perform chemiluminescent detection of DIG-labeled nucleic acids or proteins in various applications such as Northern, Southern, and Western blotting.

Automatically generated - may contain errors

Lab products found in correlation

Positively charged nylon membrane, by Merck Group (2 mentions) Nanodrop, by GE Healthcare (1 mentions) Las4000 luminescence imager, by Fujifilm (1 mentions) Pcr dig probe synthesis kit, by Merck Group (1 mentions) Dneasy extraction kit, by Qiagen (1 mentions) Nylon membrane, by Merck Group (1 mentions) Dig labeling kit, by Roche (1 mentions) Mueller hinton agar plate, by Thermo Fisher Scientific (1 mentions) Chef mapper xa system, by Bio-Rad (1 mentions) S1 nuclease, by Takara Bio (1 mentions) Cdp star chemiluminescent substrate, by Merck Group (1 mentions) Dig rna labeling kit sp6 t7, by Merck Group (1 mentions)

6 protocols using dig luminescent detection kit

1

Transgene Copy Number Determination

Check if the same lab product or an alternative is used in the 5 most similar protocols
100 mg fresh leaves transgenics were homogenized in liquid nitrogen and mixed with 500 μl CTAB buffer (2% w/v CTAB, 0.3 M NaCl, 20 mM EDTA, 100 mM Tris–Cl (pH-8) 0.2% BME). Genomic DNA was isolated according to Murray and Thompson36 . Quality and quantity of genomic DNA was checked by gel electrophoresis and nanodrop (GE, US), respectively.
Transgene copy number was checked using 10 μg of genomic DNA. DNA was digested with BamHI and separated by 0.8% agarose gel electrophoresis. DNA was blotted on Hybond-N Nylon membrane (Pharmacia) and fixed using UV cross linker (UVP HybriLinker, Analytik Jena, US). Blot was probed with hpt gene probe which was synthesized using PCR DIG probe synthesis kit (SIGMA-ALDRICH). Hybridization and detection were done using the non-radioactive method by DIG Luminescent Detection Kit as per the manufacturer guidelines (SIGMA-ALDRICH) with hybridization temperature 42 °C and washing 65 °C. Detection was carried out using CSPD solution and visualized by Fujifilm LAS4000 luminescence imager.
Sharma N., Chaudhary C, & Khurana P. (2020). Wheat Myo-inositol phosphate synthase influences plant growth and stress responses via ethylene mediated signaling. Scientific Reports, 10, 10766.
+ Open protocol
+ Expand
2

Identifying Chromosomal DNA Restriction Patterns

Check if the same lab product or an alternative is used in the 5 most similar protocols
Changes in the restriction patterns of chromosomal DNA extracted from phase variants and mutants was identified as follows. Chromosomal DNA was prepared using either a phenol extraction/caesium chloride method or a DNeasy extraction kit (Qiagen). Two micrograms of DNA was digested for 2 h at 37°C with 20–40 units of a range of methylation sensitive restriction enzymes. The resulting fragments were resolved on 1% agarose gels at 80V for 2–3 h, and visualized under UV illumination. Southern blot and hybridization (27 ) were performed using DIG-labelled (Roche) polymerase chain reaction (PCR) products as probes. These probes were generated using primers listed in Supplementary Table SI and were designed to contain one or more of the relevant restriction sites within chromosomal fragments of 0.5–10 kb. Products were visualized using a DIG luminescent detection kit (Sigma-Aldrich, UK) and autoradiography.
Anjum A., Brathwaite K.J., Aidley J., Connerton P.L., Cummings N.J., Parkhill J., Connerton I, & Bayliss C.D. (2016). Phase variation of a Type IIG restriction-modification enzyme alters site-specific methylation patterns and gene expression in Campylobacter jejuni strain NCTC11168. Nucleic Acids Research, 44(10), 4581-4594.
+ Open protocol
+ Expand
3

Northern Blot Analysis of lncAABR07025387.1

Check if the same lab product or an alternative is used in the 5 most similar protocols
Northern blotting was performed as described previously (Wang et al., 2013 (link)). Total RNA derived from H9C2 cells was separated using formaldehyde gel, transferred onto nylon membranes (Millipore, MA, United States), and hybridized with DIG-labeled probes for lncAABR07025387.1 at 60°C overnight. The lncAABR07025387.1 probe was synthesized using a DIG labeling kit (Roche, CA, United States). The membranes were then analyzed using a DIG luminescent detection kit (Sigma, MA, United States) according to the manufacturer’s protocol. A DIG-labeled U6 probe was used as an internal control.
Sun W., Wu X., Yu P., Zhang Q., Shen L., Chen J., Tong H., Fan M., Shi H, & Chen X. (2022). LncAABR07025387.1 Enhances Myocardial Ischemia/Reperfusion Injury Via miR-205/ACSL4-Mediated Ferroptosis. Frontiers in Cell and Developmental Biology, 10, 672391.
+ Open protocol
+ Expand
4

Plasmid-mediated NDM-6 gene transfer

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
As described previously (Wailan et al., 2015 (link)), filter-mating conjugation experiments were carried out using KP2e as the donor and azide-resistant E. coli J53 as the recipient. Transconjugants that possessed the blaNDM–6-carrying plasmid were detected on Mueller–Hinton agar plates (Oxoid, Hampshire, United Kingdom) containing 1 mg/L imipenem plus 100 mg/L sodium azide. The number and sizes of plasmid-located blaNDM–6 were estimated by S1 nuclease-pulsed field gel electrophoresis (S1-PFGE) followed by southern blot (Chen et al., 2015 (link)). Briefly, the linearised plasmids digested by S1 nuclease (Takara, Shiga, Japan) were separated via the CHEF Mapper XA system (Bio-Rad, Hercules, CA, United States). Furthermore, the DNA fragments were transferred to a positively charged nylon membrane (Millipore, United States) and then hybridised with a digoxigenin (DIG)-labelled probe (Roche Diagnostics, Basel, Switzerland) specific against blaNDM. The signal was detected with a DIG Luminescent Detection Kit (Sigma-Aldrich, MO, United States) and visualised through a Fusion Pulse 6 imaging system (Vilber, Marne-la-Vallée, France).
Gong Y., Lu Y., Xue D., Wei Y., Li Q., Li G., Lu S., Wang J., Wang Y., Peng Y, & Zhao Y. (2022). Emergence of a Carbapenem-Resistant Klebsiella pneumoniae Isolate Co-harbouring Dual blaNDM– 6-Carrying Plasmids in China. Frontiers in Microbiology, 13, 900831.
+ Open protocol
+ Expand
5

SARS-CoV-2 mRNA Detection by Northern Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNAs were extracted from cells infected with B.1.1or B.1.1-Akaluc and were subjected to Northern blot analysis as previously described.20 (link) In brief, a digoxigenin (DIG)-labeled random-primed probe, corresponding to 28,999 to 29,573 of the SARS-CoV-2 genomes, was generated by using a DIG RNA Labeling kit (SP6/T7) (Sigma-Aldrich), and utilized to detect viral mRNAs. The RNAs were washed with the DIG luminescent detection kit (Sigma-Aldrich) and were visualized with CDP-Star Chemiluminescent Substrate (Sigma-Aldrich), according to the manufacturer’s protocols. Bands were detected by WSE-6100LuminoGraphI (Atto).
Tamura T., Ito H., Torii S., Wang L., Suzuki R., Tsujino S., Kamiyama A., Oda Y., Tsuda M., Morioka Y., Suzuki S., Shirakawa K., Sato K., Yoshimatsu K., Matsuura Y., Iwano S., Tanaka S, & Fukuhara T. (2024). Akaluc bioluminescence offers superior sensitivity to track in vivo dynamics of SARS-CoV-2 infection. iScience, 27(5), 109647.
+ Open protocol
+ Expand
6

Northern Blot Analysis of RNA Complexes

Check if the same lab product or an alternative is used in the 5 most similar protocols
The RNA complexes were collected and run on a 15% polyacrylamide-urea gel and then transferred to positively charged nylon membranes (Millipore) followed by cross-linking through UV irradiation. The membranes were subjected to hybridization with 100 pmol 3ʹ-digoxigenin (DIG)-labeled probes overnight at 43°C, and the detection was performed using a DIG luminescent detection kit (Sigma) according to the manufacturer’s instructions.
Sun Q., Li Q, & Xie F. (2019). LncRNA-MALAT1 regulates proliferation and apoptosis of ovarian cancer cells by targeting miR-503-5p. OncoTargets and therapy, 12, 6297-6307.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!