Heat&Run gDNA removal kit

Overview

The kit contains HL-dsDNase which efficiently removes gDNA from RNA preps.

  • gDNA is removed, leaving RNA ready for reverse transcription in the same tube (Figure 1).
  • Heat-labile dsDNase can easily be inactivated without reducing the quality or quantity of RNA.
  • This procedure minimizes pipetting steps and reduces hands-on time.
  • The Heat&Run kit is especially well suited for high throughput experiments.

The high activity of the HL-dsDNase at low temperature and its heat-lability makes it ideal to use in a combination with a heat activated RT enzyme. The contaminating genomic DNA is cleave at ambient temperature and increasing the temperature over 50°C will inactivate the DNase and start the Reverse Transcriptase.

Protocol

Figure 1: Heat&Run protocol

Figure 1: Heat&Run protocol

Heat&Run causes no harmful effect on the amount of RNA being detected in subsequent RT-qPCR. In our experiments, neither mRNA, miRNA nor lnc-RNA was lost or damaged, while gDNA was efficiently and completely removed.

Figure 2: qPCR shows the Heat&Run kit removes at least 50ng of gDNA in a 10µl reaction volume.

Kit Contents

  • 10X reaction Buffer
  • HL-dsDNase (50 reactions)

Storage Conditions: Store at -20˚C.
Sample Material: The Heat&Run kit is suited for gDNA removal of RNA from any source.
Quality Control: The kit is tested for absence of RNase.

Publications

  1. Complete Coding Sequence of a Case of Chikungunya Virus Imported into Australia
    Bixing Huang, Alyssa T. Pyke, Jamie McMahon, David Warrilow.
    doi: 10.1128/genomeA.00310-17, Genome Announc. May 2017 vol. 5 no. 19 e00310-17
  2. Increased chromosomal radiosensitivity in asymptomatic carriers of a heterozygous BRCA1 mutationAnnelot Baert, Julie Depuydt, Tom Van Maerken, Bruce Poppe, Fransiska Malfait, Katrien Storm, Jenneke van den Ende, Tim Van Damme, Sylvia De Nobele, Gianpaolo Perletti, Kim De Leeneer, Kathleen B. M. Claes, and Anne Vralcorresponding
    Breast Cancer Research : BCR 18 (2016): 52. PMC. Web. 1 June 2016.
  3. Silenced vanA gene cluster on a transferable plasmid cause outbreak of vancomycin variable enterococciAudun Sivertsen, Torunn Pedersen, Kjersti Wik Larssen, Kåre Bergh, Torunn Gresdal Rønning, Andreas Radtke and Kristin Heisted
    Antimicrobial Agents and Chemotherapy, 2 May 2016, doi: 10.1128/AAC.00286-16
  4. Differentially Expressed MicroRNAs in Meningiomas Grades I and II Suggest Shared Biomarkers with Malignant Tumors
    Mohamed Raafat El-Gewely, Morten Andreassen, Mari Walquist, Anita Ursvik, Erik Knutsen, Mona Nystad, Dag H. Coucheron, Kristin Smistad Myrmel, Rune Hennig and Steinar D. Johansen
    Cancers 2016, 8(3), 31; doi:10.3390/cancers8030031
  5. Comparative Analysis of Stress Induced Gene Expression in Caenorhabditis elegans following Exposure to Environmental and Lab Reconstituted Complex Metal Mixture
    Kumar R, Pradhan A, Khan FA, Lindström P, Ragnvaldsson D, Ivarsson P, et al. (2015). PLoS ONE 10(7): e0132896. doi:10.1371/journal.pone.0132896
  6. Multi-platform assessment of transcriptome profiling using RNA-seq in the ABRF next-generation sequencing study
    Nature Biotechnology 32, 915–925 (2014) doi:10.1038/nbt.2972
    Received 14 May 2013 Accepted 01 July 2014 Published online 24 August 2014