Presentation on theme: "BiOstic™ FFPE Tissue DNA Isolation Kit (cat# 12250-50) For use with paraffin embedded tissues For use with paraffin embedded tissuesand BiOstic™ Paraffin."— Presentation transcript:
BiOstic™ FFPE Tissue DNA Isolation Kit (cat# 12250-50) For use with paraffin embedded tissues For use with paraffin embedded tissuesand BiOstic™ Paraffin Removal Reagent (cat# 12251-50) WWW.MOBIO.COM
Overview Marketing Overview R&D Data Review Sales Tools
What FFPE stands for? A formaline fixed paraffin embedded tissue (FFPE Tissue) is a piece of tissue which is first separated from a specimen animal by dissection or biopsy. Then, this tissue is fixed in order to prevent it from decaying or degeneration and to examine it clearly under a microscope for histological, pathological or cytological studies. This fixed tissue is then embedded in the wax to be cut into fine sections and stain with Hemotoxylin and Eosin Stain. After that microtoming is done, the tissue can be studied under a microscope. Fixation is the process by which the tissue is immobilized, killed and preserved for the purpose of staining and viewing it under a microscope. Fixation makes tissue permeable to staining reagents and cross- links its macromolecules so that they are stabilized and locked in position. Many fixatives are used for this purpose for example, Bouine solution, Formaline, or Liquid Nitrogen.
What is the BiOstic™ FFPE Tissue DNA Isolation Kit? A way to extract genomic DNA from formalin fixed paraffin embedded (FFPE) tissues. –used in cancer studies to view the morphology of potentially cancerous cells. –Genomic DNA used in SNP (Single nucleotide polymorphisms) genotyping, HLA (Human Leucocyte Antigen: Molecules that can identify cells as self providing the immune system to attack itself) and other genetic analysis. Why is it different? –uses a novel method to directly melt the wax away from the tissue –Does not use toxic solvents such as xylene to deparaffinize the tissue. –Yields are higher and DNA is more pure = lower Ct values and more sensitive detection.
BiOstic™ Paraffin Removal Reagent BiOstic™ Paraffin Removal Reagent is a non-hazardous, non-toxic alternative to xylene –For those who still prefer to de-wax using solvents –Can be used with the BiOstic™ FFPE Tissue DNA Isolation Kit –Cat # 12251-50
Features and Benefits Higher yields of DNA- less protocol steps results in less chances for loss of sample and higher yields of DNA More sensitive results- higher purity due to non-toxicity provides lower Ct values in real- time PCR. Safe and non-toxic- no xylene or solvents required Note: Ct value= Concentration of DNA molecules (in moles) multiplied by time.
Customer Overview Cancer research labs –Breast cancer labs HLA (Human Leukocyte Antigen) Labs Clinical research labs Who are the customers?Where do they work? How do I find them? What do they buy now? Why do they need this kit? Universities Pathology Labs Cancer institutes Large pharmaceutical companies Higher DNA recovery without toxic solvents Laboratory safety No PCR inhibition Complete kit – no extra reagents needed. Histology supplies, microscope slides, tissue stains,
Competitor Overview Competition SpecQiagen 56404 MO BIO 12250-50 Speed160 minutes140 minutes Toxic Solvent freeNoYES Buffers ready to use?NoYES Cost (US)$179$139 Price per prep (US)$3.58/prep$2.78/prep
Protocol Key steps: Digestion- tissues are melted directly in a high powered buffer that optimized proteinase K activity. 90 0 C incubation is critical for removing cross-links in DNA that inhibit PCR. Purification- DNA is isolated quickly on Spin Filters and without inhibitors.
FFPE tissue purified DNA is typically low molecular weight L MB MB Q Q DNA prepared with the MO BIO Kit shows higher molecular weight distribution as compared to the Qiagen protocol using xylene. 1 kb 500bp
High molecular weight DNA purified with the BiOstic™ FFPE Tissue DNA Kit There are two protocols available for the BiOstic FFPE Tissue kit: - Option 1: direct melting of the paraffin -Option 2: use of the non-toxic solvent BiOstic paraffin removal reagent -Both yield similar results but there is a difference in timing and personal preference.
High molecular weight DNA purified with the BiOstic™ FFPE Tissue DNA Kit 1 kb 4 kb Isolated DNA has high molecular weight species when separated with a 1.5% agarose gel L PR PR M M PR = Paraffin removal protocol M = Melt protocol
MO BIO outperformed Qiagen in qPCR results for DNA isolation from FFPE tissue DNA isolated from the BiOstic™ FFPE Tissue DNA Isolation Kit is ready for qPCR 1 CT earlier detection = 2 fold increase in sensitivity
FAQs- Technical Questions How many mgs of FFPE tissue should I use? –We tested up to 15 mg of tissue. As little as 1 slice may be used or 2-3 slices can be combined to increase yields. What factors impact the yields? –The type of tissue, length of time it was in fixative, how even the slices were made, how much tissue is in the slice, and how much sample you use. What if I want to concentrate the eluted DNA? –If you are using only 1 slice and expect yields to be in the 10ng/µl range, depending on your application, concentration may be helpful. Elute the sample in 100 µl to maximize the full recovery off of the membrane and then use standard ethanol precipitation techniques (ask technical service for our protocol). Alternatively, the UltraClean 15 Kit may be used to concentrate the sample down to a volume of 12 µl.
FAQs- Technical Questions What are expected yields? –1-3 µg from 3 sections of 5 microns is average. Measuring yield on a Nanodrop and using Picogreen will give different results depending on the amount of ssDNA present in the sample. Nanodrop will detect both ssand dsDNA and Picogreen will detect only dsDNA. –Using the MO BIO Kit with 12 mg of tissue, we have seen yields of up to 8 µg using a Nanodrop.
Spin Selling your customer – What to ask Situation, Problem, implications, need/payoffs S: Are you isolating DNA from FFPE Tissue? S: Are you currently using solvents such as xylene to deparaffinize the tissue? P: Does the protocol you are currently using has too many steps, and thus, affects the amount of yield ? P: Does the method with solvents affect the DNA purity?
SPIN Selling your customer – What to ask Situation, Problem, Implications, Need/payoffs I: The solvent wax removal method usually takes longer. How does that affect your ability to get work done? I: The use of toxic solvents may affect the quality of the DNA yield and lab safety. How does that affect your research? N: If you could use a product that would provide a safer, faster and easier way to isolate DNA from FFPE tissue, would you buy it? N: If you could use a product that would provide a gentle method to isolate DNA from FFPE tissue, and thus have more sensitive results and lower Ct values in real-time PCR due to higher DNA purity (contaminant-free), would you buy it?
Sales Tools / Promotional Activities Sales Summary and Competitive Analysis BiOstic™ FFPE Tissue DNA Isolation Kit sample: 12250-S BiOstic™ literature: MC 101 BiOstic™ pull up banner Website Press Release in January 2009 Stickers being placed in the UltraClean Tissue and Cells DNA Isolation Kits Promotion at conferences: Tri-conference, Feb 2009; AACR, April 2009; and eventually AMP 2009, October 2009.