tru XTRAC cf DNA www covaris com tru
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tru. XTRAC cf. DNA www. covaris. com
tru. XTRAC cf. DNA • Simple Workflow to Process 1 -4 m. L of Plasma • Process Plasma from EDTA or Streck Blood Collection Tubes • Magnetic bead-based purification • Highest DNA Yields enabled through Active Extraction Proprietary 2
Active Extraction by AFA – Our Key Differentiator • What: Active Extraction by AFA dissociates cf. DNA from histones and other proteins, apoptotic bodies and cellular debris • Impact: Dissociation of these components from cf. DNA helps to increase the yield and complexity of the extracted cf. DNA • Potential to see more of what you won’t see with other kits • Impact: Active Extraction ensures that the extracted cf. DNA profile is the most similar to the cf. DNA profile in plasma Proprietary 3
Why Covaris is Superior 1. 2. 3. 4. 5. cf. DNA Complexity – we uncover what other kits can’t Yield Automated purification on Thermo Kingfisher Standardization, reducing variability No genomic contamination Proprietary 4
cf. DNA Complexity – Uncover what other kits cannot • Getting the full picture of the cf. DNA profile is critical to properly interpret the patient sample • Active extraction ensures you don’t miss anything in your downstream analysis Populations of cf. DNA 1. Apoptotic Fraction • Cells undergoing apoptosis • Cells being killed by therapeutics 2. Non-apoptotic Fractions • Necrotic or other populations of cells not undergoing apoptosis • Can contain cf. DNA from cell populations containing secondary or tertiary mutations • Important for monitoring in oncology – uncover arising mutations Proprietary 5
Increased complexity of cf. DNA Higher MW fraction – important for those looking at non-apoptotic cf. DNA fraction Proprietary 6
dd. PCR – Covaris uncovers what the competition doesn’t cf. DNA extracted from matched patient samples - A-F dd. PCR targets - 1 -12 matched patient samples tru. XTRAC Competitor Competing Kit Target counts picked up by Covaris but not by a competing extraction kit You don’t know what you might be missing! Proprietary 7
Quantifying increased cf. DNA complexity • DNA integrity index (ALU 247/ALU 115) quantifies the relative amounts of higher molecular weight cf. DNA • Shear down larger fragment sizes without impacting small sizes • Provides you additional information from donor you may not normally be capturing Proprietary 8
Highly Superior Yields • • Insufficient cf. DNA yields is the single greatest issue facing most laboratories tru. XTRAC cf. DNA uses Active Extraction to achieve the highest cf. DNA yields 70% greater yields as compared to the competition Improved cf. DNA yields = Reduced input volume of plasma required • Important for pediatric, elderly, or other populations where it is challenging to draw large amounts of blood for cf. DNA extraction Proprietary 9
Yield data Higher Yield as measured by Qubit (10 donors) Higher Yield as measured by q. PCR (10 Donors) Proprietary 10
Qubit Results for K 2 EDTA Stored Samples Qubit Results EDTA Stored 10 Donors 14 12 ng Eluted 10 8 6 4 2 0 Covaris Qiagen 1 Covaris Qiagen 2 Covaris Qiagen 3 Covaris Qiagen 4 Covaris Qiagen Covaris 5 Qiagen 6 Covaris Qiagen 7 Covaris Qiagen 8 Covaris Qiagen 9 Covaris Qiagen 10 EDTA Proprietary 11 11
No Genomic Contamination Apoptotic cf. DNA Di- and tri-nucleosomal cf. DNA fractions No high MW peaks observed with Mag Bead Kit Covaris Qiagen No peak, BUT still higher MW fraction – important if looking at non-apoptotic cf. DNA fraction Proprietary 12
Standardization – Low variability • AFA provides consistent, equivalent power during active extraction • Especially important in the clinic to have consistent reproducible results • Automated workflow on the King. Fisher can reduce operator-to-operator variability Proprietary 13
Standardized Extraction Wide distribution around apoptotic fraction = Greater variability Tight distribution of apoptotic fraction Covaris Qiagen 2 1, 5 1 1 A. U. 2 0, 5 0 0, 5 50 500 -0, 5 5000 0 50 500 -0, 5 DNA size (bp) 5000 DNA size (bp) 1 Sbreast 5 -C 1 Sbreast 6 -C 1 Sbreast 7 -C 1 Sbreast 9 -C 1 Sbreast 10 -C 1 Sbreast 5 -Q 1 Sbreast 6 -Q 1 Sbreast 9 -Q 1 Sbreast 10 -Q 1 Sbreast 12 -C 1 Sbreast 13 -C 1 Sbreast 15 -C 1 Scolon 1 -C 2 Scolon 17 -C 1 Sbreast 13 -Q 1 Sbreast 14 -Q 1 Sbreast 15 -Q 1 Scolon 1 -Q 1 Scolon 2 -Q 2 Scolon 22 A-C 2 Scolon 29 A-C 2 Spancreas 25 A-C 2 Spancreas 32 A-C 1 Sbladder 1 -C 2 Scolon 17 -Q 2 Scolon 22 A-Q 2 Scolon 29 A-Q 2 Spancreas 25 A-Q 2 Spancreas 32 A-Q 1 Sbrain 1 -C 1 Slung 1 -C 1 Sbladder 1 -Q 1 Sbrain 1 -Q 1 Slung 1 -Q Proprietary 14
Improved Workflow • No pre-concentration step required • Without a preconcentration step, it is easier to integrate into mid or high throughput automation • Mag bead purification also enables high throughput automation Proprietary 15
Thermo Kingfisher – Automated purification Qubit Yields Normalized to ng DNA Eluted/ ml Plasma 9 8 ng DNA/ ml Plasma • Mid throughput automation • Demonstrated equivalence to manual processing • Process 1 -4 m. Ls of plasma • Process 6 samples at a time in a 24 -well plate on a Duo • Kingfisher Flex: Ability to process 24 samples at a time 7 6 5 4 3 2 1 0 1 ml 2 ml 3 ml King. Fisher 4 ml 1 ml Manual KP 52846 Proprietary 16
King. Fisher Processing Info and Scripts • Protocols and scripts for automated purification of cf. DNA on the King. Fisher can be found on the website here under “Technical Literature”: • http: //covaris. com/truxtrac-cfdna/#toggle-id-3 Proprietary 17
Ordering Information King. Fisher Processing Info and Scripts • Protocols and scripts for automated purification of cf. DNA on the King. Fisher can be found on the website here under “Technical Literature”: • http: //covaris. com/truxtrac-cfdna/#toggle-id-3 Proprietary 18
Additional Data Proprietary 19
Qubit Results for K 2 EDTA Stored Samples Qubit Results EDTA Stored 10 Donors 14 12 ng Eluted 10 8 6 4 2 0 Covaris Qiagen 1 Covaris Qiagen 2 Covaris Qiagen 3 Covaris Qiagen 4 Covaris Qiagen Covaris 5 Qiagen 6 Covaris Qiagen 7 Covaris Qiagen 8 Covaris Qiagen 9 Covaris Qiagen 10 EDTA Proprietary 20 20
Averaged donor yields (Qubit) Proprietary 21 21
q. PCR Results for K 2 EDTA Stored Samples Proprietary 22 22
Averaged donor yields (q. PCR) Proprietary 23 23
Yield Results for K 2 EDTA Stored Samples Proprietary 24 24
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