BiOstic™ FFPE Tissue DNA Isolation Kit (cat 12250-50)

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BiOstic FFPE Tissue DNA Isolation Kit (cat
12250-50)

• For use with paraffin embedded tissues
• and
• BiOstic Paraffin Removal Reagent
• (cat 12251-50)
• WWW.MOBIO.COM

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Overview

• Marketing Overview
• RD Data Review
• Sales Tools

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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.

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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.

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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

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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.

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Customer Overview
Who are the customers?
Where do they work?

• Cancer research labs
• Breast cancer labs
• HLA (Human Leukocyte Antigen) Labs
• Clinical research labs

• Universities
• Pathology Labs
• Cancer institutes
• Large pharmaceutical companies

How do I find them? What do they buy now?
Why do they need this kit?

• Higher DNA recovery without toxic solvents
• Laboratory safety
• No PCR inhibition
• Complete kit no extra reagents needed.

• Histology supplies, microscope slides, tissue
  stains,

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Competitor Overview
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RD Data
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Protocol
Key steps Digestion- tissues are melted
directly in a high powered buffer that optimized
proteinase K activity. 900C incubation is
critical for removing cross-links in DNA that
inhibit PCR. Purification- DNA is isolated
quickly on Spin Filters and without inhibitors.
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FFPE tissue purified DNA is typically low
molecular weight
DNA prepared with the MO BIO Kit shows higher
molecular weight distribution as compared to the
Qiagen protocol using xylene.
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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.

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High molecular weight DNA purified with the
BiOstic FFPE Tissue DNA Kit
L PR PR M M
PR Paraffin removal protocol M Melt protocol
Isolated DNA has high molecular weight species
when separated with a 1.5 agarose gel
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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
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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.

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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.

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Sales Tools
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Spin Selling your customer What to
askSituation, 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?

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SPIN Selling your customer What to
askSituation, 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?

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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.

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Ordering information and Contacts

• Cat12250-50 (50 preps) and 12250-S (sample size)
• RD Suzanne kennedy, skennedy_at_mobio.com
• Technical Support Nik von Atzigen,
  nvonatzigen_at_mobio.com
• Marketing- Peggy Maunaury, pmaunaury_at_mobio.com