Heat&Run gDNA removal kit

Protocol

The kit contains HL-dsDNase which efficiently removes gDNA from RNA preps before running RT-qPCR.

  • gDNA is removed, leaving RNA ready for reverse transcription in the same tube (Figure 1).
  • Heat-labile dsDNase can easily be inactivated.
  • 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 cleaved at ambient temperature and increasing the temperature over 50°C will inactivate the DNase and start the Reverse Transcriptase.

Do you require gDNA removal in applications other than RT-qPCR? Contact our support team for assistance in implementing dsDNase treatment in your workflow.

Image

Figure 1: Heat&Run protocol

Learn more about dsDNase
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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 in qPCR settings from samples with good RNA quality as some loss of RNA integrity can be expected.
Quality Control: The kit is tested for absence of RNase.

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Publications

Heat&Run®gDNA Removal Kit Applications

  1. Metagenomic analysis of thevirome of mosquito excreta.
    Ramírez AL, Colmant AMG, Warrilow D, Huang B, Pyke AT, McMahon JL, Meyer DB, Graham RMA, Jennison AV, Ritchie SA, van den Hurk AF.
    mSphere. 2020; 5(5): e00587-20.

  2. Long non-coding RNA RAMS11 promotes metastatic colorectal cancer progression.
    Silva-Fisher JM, Dang HX, White NM, Strand MS, Krasnick BA, Rozycki EB, Jeffers GGL, Grossman JG, Highkin MK, Tang C, Cabanski CR, Eteleeb A, Mudd J, Goedegebuure SP, Luo J, Mardis ER, Wilson RK, Ley TJ, Lockhart AC, Fields RC, Maher CA.
    Nat Commun. 2020; 11, 2156.

  3. Genome sequences of chikungunya virus strains from Bangladesh and Thailand.
    Pyke AT, McMahon J, Burtonclay P, Nair N, De Jong A.
    Microbiol Resour Announc. 2020; 9 (2): e01452-19.

  4. Performance assessment of total RNA sequencing of human biofluids and extracellular vesicles. 
    Everaert C, Helsmoortel H, Decock A.
    Scientific Reports. 2019; 9, 17574.

  5. Illumina sequencing of clinical samples for virus detection in a public health laboratory.
    Huang B, Jennison A, Whiley D.
    Scientific Reports. 2019; 9, 5409.

  6. CRISPR Interference for Rapid Knockdown of Essential Cell Cycle Genes in Lactobacillus plantarum.
    Myrbråten IS, Will K, Salehian Z, Håvarstein LS, Straume D, Mathiesen G, Kjos M.
    mSphere. 2019; 4 (2): e00007-19.

  7. First complete genome sequences of dengue virus serotype 2 strains from the Solomon Islands and Vanuatu.
    Pyke AT, Cameron J, McMahon J, De Jong A, Burtonclay P.
    Microbiology Resource Announcements. 2019; 8(2): e01360-18.

  8. Antisense Oligonucleotide-Based Downregulation of the G56R Pathogenic Variant Causing NR2E3-Associated Autosomal Dominant Retinitis Pigmentosa. 
    Naessens S, Ruysschaert L, Lefever S, Coppieters F, De Baere E.
    Genes. 2019; 10 (5): 363.

  9. The N‐terminal p.(Ser38Cys) TIMP3 mutation underlying Sorsby fundus dystrophy is a founder mutation disrupting an intramolecular disulfide bond.
    Naessens S, De Zaeytijd J, Syx D, Vandebroucke RE, Smeets F, Van Cauwenbergh C, Leroy BP, Peelman F, Coppieters F.
    Human Mutation. 2019; 40 (5): 539-551.

  10. Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes.
    Loontiens S, Depestel L, Vanhauwaert S, Dewyn G, Gistelinck C, Verboom K, Van Loocke W, Matthijssens F, Willaert A, Vandesompele J, Speleman F, Durinck K.
    BMC Genomics. 2019; 20, 228.

  11. The validation of Short Interspersed Nuclear Elements (SINEs) as a RT-qPCR normalization strategy in a rodent model for temporal lobe epilepsy.
    Crans RAJ, Janssens J, Daelemans S, Wouters E, Raedt R, Van Dam D, De Deyn PP, Van Craenenbroeck K, Stove CP.
    PLoS ONE. 2019; 14 (1): e0210567.

  12. Resistance Training Alone or Combined With N-3 PUFA-Rich Diet in Older Women: Effects on Muscle Fiber Hypertrophy.
    Strandberg E, Ponsot E, Piehl-Aulin K, Falk G, Kadi F.
    The Journals of Gerontology. 2019; A (74) 4: 489-494.

  13. Switching-Off Adora2b in Vascular Smooth Muscle Cells Halts the Development of Pulmonary Hypertension.
    Mertens TCJ, Hanmandlu A, Tu L, Phan C, Collum SD, Chen NY, Weng T, Davies J, Liu C, Eltzschig HK, Jyothula SSK, Rajagopal K, Xia Y, Guha A, Bruckner BA, Blackburn MR, Guignabert C, Karmouty-Quintana H.
    Front Physiol. 2018; 9:555.

  14. Enterococcus faecium TIR-Domain Genes Are Part of a Gene Cluster Which Promotes Bacterial Survival in Blood.
    Wagner TM, Janice J, Paganelli FL, Willems RJ, Askarian F, Pedersen T, Top J, de Haas C, van Strijp JA, Johannessen M, Hegstad K.
    Int. Journal of Microbiology. 2018; 1435820.

  15. Comparative Analysis of the Nodule Transcriptomes of Ceanothus thyrsiflorus (Rhamnaceae, Rosales) and Datisca glomerata (Datiscaceae, Cucurbitales).
    Salgado MG, van Velzen R, Van Nguyen T, Battenberg K, Berry AM, Lundin D, Pawloski K.
    Frontiers in Plant Science. 2018; 9: 1629.

  16. New genotypes of Liao ning virus (LNV) in Australia exhibit an insect-specific phenotype.
    Prow NA, Mah MG, Deerain JM, Warrilow D, Colmant AMG, O'Brien CA, Harrison JJ, McLean BJ, Hewlett EK, Piyasena TBH, Hall-Mendelin S, van den Hurk A, Watterson D, Huang B, Schulz BL, Webb CE, Johansen CA, Chow WK, Hobson-Peters J, Cazier C, Coffey LL, Faddy HM, Suhrbier A, Bielefeldt-Ohmann H, Hall RA.
    Journal of General Virology. 2018; 99: 596-609.

  17. Expressed repetitive elements are broadly applicable reference targets for normalization of reverse transcription-qPCR data in mice.
    Renard M, Vanhauwaert S, Vanhomwegen M, Rihani A, Vandamme N, Goossens S, Berx G, Van Vlierberghe P, Haigh JJ, Decaesteker B, Van Laere J, Lambertz I, Speleman F, Vandesompele J, Willaert A.
    Scientific Reports. 2018; 8: 7642.

  18. Allene oxide synthase, allene oxide cyclase and jasmonic acid levels in Lotus japonicus nodules.
    Zdyb A, Salgado MG, Demchenko KN, Brenner WG, Płaszczyca M, Stumpe M, Herrfurth C, Feussner I, Pawlowski K.
    PLoS ONE. 2018; 13(1): e0190884.

  19. Development of Escherichia coli‐based gene expression profiling of sewage sludge leachates.
    Goswami M, Khan FA, Ibrisevic A, Olsson P, Jass J.
    J Appl Microbiol. 2018; 125 (5): 1502-1517.

  20. Transcriptome Profiling Reveals Interplay of Multifaceted Stress Response in Escherichia coli on Exposure to Glutathione and Ciprofloxacin.
    Goswami M, Narayana Rao AVSS.
    mSystems. 2018; 13;3(1): e00001-18.

  21. Food Spoilage-Associated Leuconostoc, Lactococcus, and Lactobacillus Species Display Different Survival Strategies in Response to Competition
    Andreevskaya M, Jääskeläinen E, Johansson P, Ylinen A, Paulin L, Björkroth J, Auvinen P.
    Appl. Environ. Microbiol. 2018; 84 (13): e00554-18.

  22. Emerging recombinant noroviruses identified by clinical and waste water screening.
    Lun JH, Hewitt J, Sitabkhan A, Eden JS, Enosi Tuipulotu D, Netzler NE, Morrell L, Merif J, Jones R, Huang B, Warrilow D, Ressler KA, Ferson MJ, Dwyer DE, Kok J, Rawlinson WD, Deere D, Crosbie ND, White PA.
    Emerg Microbes Infect. 2018; 7 (1): 50.

  23. Complete Coding Sequence of a Case of Chikungunya Virus Imported into Australia.
    Huang B, Pyke AT, McMahon J, Warrilow D.
    Genome Announc. 2017; 5 (19): e00310-17.

  24. Evaluation of relative quantification of alternatively spliced transcripts using droplet digital PCR.
    Van Heetvelde, M, Van Loocke W, Trypsteen W, Baert A, Vanderheyden K, Crombez B, Vandesompele J, De Leeneer K, Claes KBM.
    Biomol Detect Quantif. 2017; 13, 40-48.

  25. Reference genes for gene expression studies in the mouse heart.
    Ruiz-Villalba A, Mattiotti A, Gunst QD, Cano-Ballesteros S, van den Hoff MJ, Ruijter JM.
    Scientific Reports. 2017; 7 (1): 24.

  26. Increased chromosomal radiosensitivity in asymptomatic carriers of a heterozygous BRCA1 mutation.
    Baert A, Depuydt J, Van Maerken T, Poppe B, Malfait F, Storm K, van den Ende J, Van Damme T, De Nobele S, Perletti G, De Leeneer K, Claes KB, Vral A.
    Breast Cancer Research. 2016; 18 (1): 52.

  27. Silenced vanA Gene Cluster on a Transferable Plasmid Caused an Outbreak of Vancomycin-Variable Enterococci.
    Sivertsen A, Pedersen T, Larssen KW, Bergh K, Rønning TG, Radtke A, Heisted K.
    Antimicrobial Agents and Chemotherapy. 2016; 60 (7): 4119-4127.

  28. Differentially Expressed MicroRNAs in Meningiomas Grades I and II Suggest Shared Biomarkers with Malignant Tumors.
    El-Gewely MR, Andreassen M, Walquist M, Ursvik A, Knutsen E, Nystad M, Coucheron DH, Myrmel KS, Hennig R, Johansen SD.
    Cancers 2016, 8(3), 31.

  29. 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, Olsson PE, Jass J.
    PLoS ONE. 2015; 10(7): e0132896.

  30. Multi-platform assessment of transcriptome profiling using RNA-seq in the ABRF next-generation sequencing study.
    Li S, Tighe SW, Nicolet CM, Grove D, Levy S, Farmerie W, Viale A, Wright C, Schweitzer PA, Gao Y, Kim D, Boland J, Hicks B, Kim R, Chhangawala S, Jafari N, Raghavachari N, Gandara J, Garcia-Reyero N, Hendrickson C, Roberson D, Rosenfeld JA, Smith T, Underwood JG, Wang M, Zumbo P, Baldwin DA, Grills GS, Mason CE.
    Nature Biotechnology. 2014; 32, 915–925.