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BisulFlash DNA Modification Kit


For lightning fast bisulfite treatment of DNA for methylation-specific PCR analysis

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Suggested Workflow
DNA Isolation
DNA Bisulfite Conversion
PCR Analysis
Complete Cytosine Conversion: 200 ng of genomic DNA isolated from 3 cancer cell lines was treated with the BisulFlash™ DNA Modification Kit. Next, the unconverted and converted DNA in each treated sample were determined using unconverted DNA-specific and converted DNA-specific primers (β-actin, 110 bps), respectively. A: real time PCR; B: end-point PCR. The BisulFlash™ kit treated DNA was completely converted, and no unconverted DNA in the treated samples was determined after 45 cycles.
Effective DNA Protection: Fully methylated human genomic DNA at various amounts (0.2 ng-200 ng) were converted using the BisulFlash™ DNA Modification Kit. 1
High accuracy of DNA conversion is achieved by the BisulFlash™ DNA Modification Kit. 50 ng of genomic DNA methylated in all CpG sites by DNA methylases was treated with the BisulFlash™ DNA Modification Kit. Converted DNA was amplified by real time qPCR using primers for multiple promoters containing numerous CpG sites and then directly sequenced.
Schematic procedure of the BisulFlash™ DNA Modification Kit to obtain converted DNA.
Input Type: DNA
Research Area: DNA Methylation
Target Application: Sample Modification
Vessel Format: Columns/Tubes
100% Guarantee: 6 months
Catalog No.SizePriceQty
P-1026-05050 reactions $122.00 
Availability: Usually Ships in 1 Day or Same day delivery Same Day NY Delivery 
Product Overview

The BisulFlash™ DNA Modification Kit is a complete set of optimized buffers and reagents to perform DNA modification using an expedited DNA bisulfite conversion technology developed by Epigentek, geared specifically for methylation-specific PCR (MSP) as well as post-bisulfite based NGS applications. Through a proprietary composition which allows DNA denaturation and bisulfite conversion to be processed at the same time, the complete procedure is reduced to only 30 minutes. Furthermore, it prevents more than 90% of DNA loss, completely converting unmethylated cytosine into uracil.

Perfecting Bisulfite Conversion
Traditional methods involve a separate denaturation step followed by a subsequent sodium bisulfite DNA conversion step -- but with the BisulFlash™ method, DNA denaturation status is concurrently sustained throughout the entire DNA bisulfite conversion process. This breakthrough approach enables the DNA conversion process to be significantly faster with higher conversion efficiency and accuracy. We continue to innovate with the development of the new BisulFlash™ kit by identifying four critical components of bisulfite conversion:
  • Speed: Reduce the entire procedure to as short as 30 minutes without any reagent setup time.
  • Efficiency: Completely convert unmethylated cytosine into uracil -- modified DNA > 99.9%.
  • DNA Protection: Protect against DNA degradation of which more than 90% of DNA loss can be prevented, allowing for greater recovery.
  • Sensitivity: Start with the lowest amount of input DNA for modification -- only 0.2 ng or just 50 cells.

The convenient ready-to-use, DNA conversion mix solution and single temperature incubation along with the features mentioned above allow for true perfection in bisulfite conversion. The BisulFlash™ DNA Modification Kit is suitable for MS-PCR/MSP and real time MSP.

Bisulfite Sequencing Option
The BisulFlash™ kit includes a special step made specifically for better sequencing results and is 100% compatible with all Illumina® platforms. For ideal NGS applications, we recommend combining this technique with our EpiNext Post-Bisulfite Library Preparation Kit.

Fig. 1. High accuracy of DNA conversion is achieved by the BisulFlash™ DNA Modification Kit. 50 ng of genomic DNA methylated in all CpG sites by DNA methylases was treated with the BisulFlash™ DNA Modification Kit. Converted DNA was amplified by real time qPCR using primers for multiple promoters containing numerous CpG sites and then directly sequenced.

Comparative Overview of Commercial Kits
Data was obtained through actual use of the kit, customer feedback, or information provided by the supplier's datasheet or website.

  BisulFlash Supplier 1 Supplier 2 Supplier 3
Processing Time 30 min >3 hours >6 hours 12-16 hours
Correct Conversion 99.9% 99.5% 99.4% 98%
Error Conversion <0.1% >0.3% N/A >1.5%
Correct/Error Conversion Ratio ≅ 1000 ≅ 330 N/A ≅ 65
DNA Degradation Very low Medium Low High
Minimum Starting DNA 0.2 ng 0.5 ng 1 ng 1 ng
Convenience Very high High Medium Low
Product Details

Principle & Procedure
As a next generation bisulfite conversion tool, the BisulFlash™ DNA Modification Kit contains all reagents required for an ultra-fast bisulfite conversion on a DNA sample. With the ready-to-use conversion mix solution, DNA denaturation status is sustained throughout the entire bisulfite DNA conversion process. This proprietary solution allows the bisulfite reagents to rapidly convert all cytosine to uracil with negligible methylcytosine conversion. The unique DNA protection reagents contained in the mix can prevent the chemical and thermophilic degradation of DNA in the bisulfite treatment. The non-toxic DNA capture solution enables DNA to tightly bind to the column filter, thus DNA cleaning can be carried out on the column to effectively remove residual bisulfite and salts.

Rapid Results
Only 30 minutes are required for the entire BisulFlash™ procedure -- from converting your sample DNA to pure bisulfite modified DNA. That is significantly faster than any currently used kits (2-8 h) or homebrew methods (>16 h). The BisulFlash™ kit provides everything required for a successful bisulfite conversion and DNA cleanup in the shortest time with the fewest steps possible.

Perfect DNA Conversion
Each reaction with the BisulFlash™ kit can use 0.2 ng – 1 µg of DNA. For optimal conversion, the DNA amount is 50-200 ng. The novel procedure and proprietary ready-to-use DNA conversion mix solution allow DNA denaturation and bisulfite conversion to occur at the same time and enable all cytosines to be converted to uracil (>99.9%), while 5-methylcytosine remains the same. The highly efficient cytosine conversion is proven by the low CT values obtained when amplifying converted DNA using real-time PCR (Fig. 2). This perfect conversion rate is superior to other bisulfite kits available on the market and provides repeatable and dependable downstream analysis.

Powerful DNA Protection
DNA protection reagents are added into the DNA conversion mix solution to prevent DNA from chemical and thermophilic degradation in the bisulfite treatment and provide effective DNA denaturation, resulting in single-stranded DNA necessary for complete cytosine conversion. The prevention of DNA degradation enables subsequent amplification and analysis of large PCR fragments (Fig. 3). The efficient integrated DNA cleanup and unique elution buffer allow for long-term storage (> 6 months) and multiple freezing/thawing of the converted DNA without affecting the DNA quality.

Fig. 3. Complete Cytosine Conversion: 200 ng of genomic DNA isolated from 3 cancer cell lines was treated with the BisulFlash™ DNA Modification Kit. Next, the unconverted and converted DNA in each treated sample were determined using unconverted DNA-specific and converted DNA-specific primers (β-actin, 110 bps), respectively. A: real time PCR; B: end-point PCR. The BisulFlash™ kit treated DNA was completely converted, and no unconverted DNA in the treated samples was determined after 45 cycles.

Fig. 4. Effective DNA Protection: Fully methylated human genomic DNA at various amounts (0.2 ng-200 ng) were converted using the BisulFlash™ DNA Modification Kit. 1 µl of 20 µl eluate was used for real time qPCR and a pair of primers was used to amplify converted DNA. As little as 0.2 ng DNA is sufficient for bisulfite conversion using the BisulFlash™ DNA Modification Kit. A: real time PCR; B: starting DNA amount-CT value curve.
Product Components

BF1 (Conversion Mix Solution)
BF2 (Capture Solution)
BF3 (Desulphonation Solution)
BF4 (Elution Solution)
BF5 (Conversion Enhancer)
BF6 (Denaturation Enhancer)
F-Spin Columns
F-Collection Tubes
User Guide

User Guide & MSDS

[User Guide]*
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

Product Citations

Haraguchi K et. al. (September 2017). The methylation status and expression of human telomerase reverse transcriptase is significantly high in oral carcinogenesis. APMIS. 125(9):797-807.

Zhang D et. al. (December 2016). Non-CpG methylation by DNMT3B facilitates REST binding and gene silencing in developing mouse hearts. Nucleic Acids Res.

Ejima-Yamada K et. al. (November 2016). Epstein-Barr virus infection and gene promoter hypermethylation in rheumatoid arthritis patients with methotrexate-associated B cell lymphoproliferative disorders. Virchows Arch.

Yang Y et. al. (September 2016). SAC3B, a central component of the mRNA export complex TREX-2, is required for prevention of epigenetic gene silencing in Arabidopsis. Nucleic Acids Res.

Jiang Y et. al. (September 2016). Trichloroethylene-Induced DNA Methylation Changes in Male F344 Rat Liver. Chem Res Toxicol.

Sun LX et. al. (August 2016). Global DNA Methylation Changes in Nile Tilapia Gonads during High Temperature-Induced Masculinization. PLoS One. 11(8):e0158483.

Kaur M et. al. (July 2016). Development of a multiplex MethyLight assay for the detection of DAPK1 and SOX1 methylation in epithelial ovarian cancer in a north Indian population. Genes Genet Syst.

Li Y et. al. (July 2016). Mir-449a, a potential diagnostic biomarker for WNT group of medulloblastoma. J Neurooncol.

Lin L et. al. (June 2016). Epigenetic regulation of reelin expression in multiple myeloma. Hematol Oncol.

Wang X et. al. (May 2016). The cytosolic Fe-S cluster assembly component MET18 is required for the full enzymatic activity of ROS1 in active DNA demethylation. Sci Rep. 6:26443.

Zuberi M et. al. (April 2016). Utility of Serum miR-125b as a Diagnostic and Prognostic Indicator and Its Alliance with a Panel of Tumor Suppressor Genes in Epithelial Ovarian Cancer. PLoS One. 11(4):e0153902.

Li Y et. al. (March 2016). Aberrant Methylation of the E-Cadherin Gene Promoter Region in Endometrium and Ovarian Endometriotic Cysts of Patients with Ovarian Endometriosis. Gynecol Obstet Invest.

Amacher DE et. al. (March 2016). A 2015 survey of established or potential epigenetic biomarkers for the accurate detection of human cancers. Biomarkers. :1-17.

Abel S et. al. (March 2016). Plasmodium yoelii infection of BALB/c mice results in expansion rather than induction of CD4<sup>+</sup> Foxp3<sup>+</sup> regulatory T cells. Immunology.

Chen C et. al. (March 2016). Epigenetic modification of PKMζ rescues aging-related cognitive impairment. Sci Rep. 6:22096.

Yang DL et. al. (December 2015). Dicer-independent RNA-directed DNA methylation in Arabidopsis. Cell Res.

Li H et. al. (November 2015). Genetic analysis of the clonal stability of Chinese hamster ovary cells for recombinant protein production. Mol Biosyst.

Wang W et. al. (October 2015). Promoter hypermethylation of PTPL1, PTPN6, DAPK, p16 and 5-azacitidine inhibits growth in DLBCL. Oncol Rep.

Felli N et. al. (October 2015). AP2α controls the dynamic balance between miR-126&amp;126* and miR-221&amp;222 during melanoma progression. Oncogene.

Wang D et. al. (September 2015). Hypermethylation of the Keap1 gene inactivates its function, promotes Nrf2 nuclear accumulation, and is involved in arsenite-induced human keratinocyte transformation. Free Radic Biol Med.

Shao LW et. al. (September 2015). ATRX loss in adult supratentorial diffuse astrocytomas correlates with p53 over expression and IDH1 mutation and predicts better outcome in p53 accumulated patients. Histol Histopathol. :11664.

Barrett HL et. al. (September 2015). Placental lipase expression in pregnancies complicated by preeclampsia: a case-control study. Reprod Biol Endocrinol. 13(1):100.

Zhang S et. al. (July 2015). The g.-165 T>C Rather than Methylation Is Associated with Semen Motility in Chinese Holstein Bulls by Regulating the Transcriptional Activity of the HIBADH Gene. PLoS One. 10(7):e0127670.

Tatura R et. al. (June 2015). Relevance of Foxp3+ regulatory T cells for early and late phases of murine sepsis. Immunology.

Lei M et. al. (March 2015). Regulatory link between DNA methylation and active demethylation in Arabidopsis. Proc Natl Acad Sci U S A. 112(11):3553-7.

Zhang W et. al. (March 2015). Correlation between the expression of DNMT1, and GSTP1 and APC, and the methylation status of GSTP1 and APC in association with their clinical significance in prostate cancer. Mol Med Rep.

Ito M et. al. (February 2015). Association of Fusobacterium nucleatum with clinical and molecular features in colorectal serrated pathway. Int J Cancer.

Zuberi M et. al. (January 2015). RASSF1 & PTEN promoter hypermethylation influences the outcome in epithelial ovarian Cancer Clini Ovarian Other Gyn Cancer.

Nosho K et. al. (January 2015). Clinicopathological and molecular characteristics of serrated lesions in Japanese elderly patients. Digestion. 91(1):57-63.

Zheng L et. al. (January 2015). FOXA1 positively regulates gene expression by changing gene methylation status in human breast cancer MCF-7 cells. Int J Clin Exp Pathol. 8(1):96-106.

Arechederra M et. al. (December 2014). p38 MAPK down-regulates fibulin 3 expression through methylation of gene regulatory sequences. Role in migration and invasion. J Biol Chem.

Erdem B et. al. (October 2014). Promoter hypermethylation of p16 and APC in gastrointestinal cancer patients. Turk J Gastroenterol. 25(5):512-7.

Minning C et. al. (August 2014). Exploring breast carcinogenesis through integrative genomics and epigenomics analyses. Int J Oncol.

Naito T et. al. (August 2014). IGF2 differentially methylated region hypomethylation in relation to pathological and molecular features of serrated lesions. World J Gastroenterol. 20(29):10050-61.

Yamamuro C et. al. (June 2014). Overproduction of stomatal lineage cells in Arabidopsis mutants defective in active DNA demethylation. Nat Commun. 5:4062.

Zhang DH et. al. (May 2014). High-risk human papillomavirus infection associated with telomere elongation in patients with esophageal squamous cell carcinoma with poor prognosis. Cancer.

Wei D et. al. (May 2014). Increased DNA Methyltransferase 3b (Dnmt3b) -mediated CpG Island Methylation Stimulated by Oxidative Stress Inhibits Expression of a Gene Required for Neural Tube and Neural Crest Development in Diabetic Pregnancy. Diabetes.

Ito M et. al. (April 2014). MicroRNA-31 expression in relation to BRAF mutation, CpG island methylation and colorectal continuum in serrated lesions. Int J Cancer.

Hadoux J et. al. (April 2014). SDHB mutations are associated with response to temozolomide in patients with metastatic pheochromocytoma or paraganglioma. Int J Cancer.

Basu A et. al. (April 2014). The CpG Island Encompassing the Promoter and First Exon of Human DNMT3L Gene Is a PcG/TrX Response Element (PRE). PLoS One. 9(4):e93561.

Guang W et. al. (February 2014). Genetic regulation of MUC1 expression by Helicobacter pylori in gastric cancer cells. Biochem Biophys Res Commun. 445(1):145-50.

Saheb A et. al. (February 2014). Probing for DNA methylation with a voltammetric DNA detector. Analyst. 139(4):786-92.

Guo F et. al. (February 2014). Alternative splicing, promoter methylation, and functional SNPs of sperm flagella 2 gene in testis and mature spermatozoa of Holstein bulls. Reproduction. 147(2):241-52.

Kehrmann J et. al. (January 2014). Impact of 5-Aza-2`-deoxycytidine and Epigallocatechin-3-gallate for induction of human regulatory T cells. Immunology.

Zhang D et. al. (November 2013). Homocysteine-related hTERT DNA demethylation contributes to shortened leukocyte telomere length in atherosclerosis. Atherosclerosis. 231(1):173-9.

Wang J et. al. (April 2013). The role of EZH2 and DNA methylation in hMLH1 silencing in epithelial ovarian cancer. Biochem Biophys Res Commun. 433(4):470-6.

Sato A et. al. (February 2013). CCAAT/enhancer-binding protein-α suppresses lung tumor development in mice through the p38α MAP kinase pathway. PLoS One. 8(2):e57013.

Kim WG et. al. (January 2013). Reactivation of the silenced thyroid hormone receptor β gene expression delays thyroid tumor progression. Endocrinology. 154(1):25-35.

Founds SA et. al. (August 2012). Variations in discovery-based preeclampsia candidate genes. Clin Transl Sci. 5(4):333-9.

Qian W et. al. (June 2012). A histone acetyltransferase regulates active DNA demethylation in Arabidopsis. Science. 336(6087):1445-8.

Martínez-Macías MI et. al. (February 2012). A DNA 3' phosphatase functions in active DNA demethylation in Arabidopsis. Mol Cell. 45(3):357-70.

Yang J et. al. (January 2012). Stem cell gene SALL4 suppresses transcription through recruitment of DNA methyltransferases. J Biol Chem. 287(3):1996-2005.

Yang X et. al. (October 2011). Neurodegenerative and inflammatory pathway components linked to TNF-α/TNFR1 signaling in the glaucomatous human retina. Invest Ophthalmol Vis Sci. 52(11):8442-54.

Komori HK et. al. (October 2011). Application of microdroplet PCR for large-scale targeted bisulfite sequencing. Genome Res. 21(10):1738-45.

Geyer KK et. al. (August 2011). Cytosine methylation regulates oviposition in the pathogenic blood fluke Schistosoma mansoni. Nat Commun. 2:424.

Carrard A et. al. (August 2011). Increased DNA methylation status of the serotonin receptor 5HTR1A gene promoter in schizophrenia and bipolar disorder. J Affect Disord. 132(3):450-3.

Cho YH et. al. (July 2011). Identification of transcriptional regulatory elements required for the Mup2 expression in circadian clock mutant mice. Biochem Biophys Res Commun. 410(4):834-40.

Kumar Suresh et. al. Salt-Induced Tissue-Specific Cytosine Methylation Downregulates Expression of HKT Genes in Contrasting Wheat (Triticum aestivum L.) Genotypes DNA and Cell Biology. Feb. 2017

Zhaofei Fan et. al. Significant association of cyp19a promoter methylation with environmental factors and gonadal differentiation in olive flounder Paralichthys olivaceus CBP Part A: Molecular & Integrative Physiology. 204(Feb. 2017)

Customer Reviews

Rating by c********@niehs.nih.gov Verified Purchase Reviewed on: Tuesday 12 October, 2010
Application Description
Pyrosequencing from blood samples

Other Thoughts
Kit was really good for higher starting amounts, not as good as others for lower amounts. If starting quantity not limiting, good kit to use.
Rating by m*****@oakland.edu Verified Purchase Reviewed on: Tuesday 12 October, 2010
Application Description
We have used your kit to make converted DNA from immature and mature neural retina to do Methylation specific PCR of the Rhodopsin gene. (off in immature, and on in mature) We can successfully confirm hypomethylation changes using DNA we processed with your kit. I have thus mentioned your kit specifically in an NIH grant application I just submitted last Monday. (R01).

Procedural Details
We did not that while a range of DNA can be processed with your kit, in reality the upper end of DNA range is best and often necessary. So processing 1 ug of DNA per treatment/column, is best to ensure that qPCR detection works well. Amounts lower than 500 ng would often fail for qPCR unless we did a precip and concentration. 1 ug input into the treatment ensures no need to reconcentrate treated DNA and successful qPCR with 2 ul per qPCR reaction. We used Amplitaq gold, gold buffer, and sybr green we add ourselves in an MX3000 qPCR unit.

Other Thoughts
Methylation primers and un-methylated primer reactions each support each other. (methyl decreases as unmethy increases). Differences between ON and OFF gene state were well in excess of 10 fold, making the differences quite clear to us. We also know what to expect from previous qChIP analysis of 5-methylcytosine.
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