Investigator 24plex GO! · Blood on FTA cards STAR Q Punch AS: High -throughput punching and assay...

Sample to Insight

High Throughput Solutions for DNA Databasing

Keith Elliott

Sample to Insight

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Section A (required)

Section B (recommended)

Consensus

Regional consensus

SE33

D2S441D10S1248D22S1045

D1S1656

D12S391TPOXD7S820CSF1POD5S818D13S317D3S1358D8S1179D21S11vWAD18S51FGA

D16S539TH01

D2S1338D19S433DYS391Amelogenin

Investigator 24plex GO! Kit

Improving global STR analysis - Investigator 24plex QS Kits 2

24plex

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CODIS13

13 15

ESS

12 15

CODISexpsn

19 22

Sample to Insight

Kit concept and overview


For reference and casework samples after DNA purification

For highest flexibility and best possible results with difficult samples

For reference samples from FTA, BB or swabs

For fast and cost-efficient results

Investigator 24plex GO!Investigator 24plex QS

Investigator 24plex Kit family

Powerful portfolio for the expanded CODIS set of markers, tailored to optimizing process efficiency for any application

Sample to Insight

Investigator 24plex GO! Kit — direct amp workflows


Buccalswabs

5 min Fast Swab Lysis protocolInvestigatior STR GO! Lysis Buffer

Prepare PCR master mix

PCR cycling (27 cycles)

Add 2 µl swab lysate

Buccalcells

on FTA®

card


Prepare PCR master mixInvestigator STR GO! Punch Buffer

1.25 mm punchfrom the centre

Add punch

Blood on FTA® card

1.25 mm punchfrom the centre


Prepare PCR master mix

Add punch

Sample to Insight

Investigator 24plex GO! Kit — direct amp workflow


Preparation of PCR Master Mix

• Investigator STR GO! Punch Buffer (200/1000), Cat No.: 386526/386528• Investigator STR GO! Lysis Buffer (200), Cat No.: 386516

Component Blood on FTA or other paper

Buccal cells on FTA cards or other paper Buccal swabs

Fast Reaction Mix 2.0 7.5 µl 7.5 µl 7.5 µl

Primer Mix 12.5 µl 12.5 µl 12.5 µl

Punch Buffer - 2.0 µl -

Total volume 20 µl 22 µl 20 µl

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Blood on FTA paper


Profile example Investigator 24plex GO! Kit

Single 1.2 mm punch

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Buccal cells on FTA paper



Single 1.2 mm punch (EasiCollect®)

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Buccal cells on swab



1 swab lysed in 500 µl Investigator GO! Lysis buffer in 5 minutes, 2 µl as template

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Investigator Quality Sensor


Internal Performance Control – more information for better results

• The kits include a Quality Sensor that will provide you with more information about a profile to improve your workflow and save unnecessary re-runs of a sample

• The internal Quality Sensor is included in the Primer Mix and amplified simultaneously with the STR markers

• The Quality Sensor gives one small (appr. 70 bp) and one large (435 bp) amplicon• You will be able to distinguish: ◦ Successful amplification◦ No DNA ◦ Failed PCR amplification◦ Inhibited DNA◦ Degraded DNA

The Quality Sensor provides additional information to improve quality of results and reduce costs

Sample to Insight

24plex Quality Sensor explained


Normal non-degraded / non-inhibited sample

No DNA

Failed PCR Amplification

The Quality Sensor confirms that your sample is not degraded or inhibited

QS confirms a successful PCR without DNA. Re-run will not lead to better results

QS confirms failed PCR. Re-run will most likely lead to better results

Sample to Insight

24plex Quality Sensor explained


Inhibited DNA sample

Degraded DNA

QS indicates a degraded sample. Re-run will not improve profile. Investigator 24plex already gives the best results at standard setup

QS indicates PCR inhibition. An inhibited profile may be rescued by re-running the PCR with a higher sample dilution

Sample to Insight

Hamilton easyPunch

PowerPoint Style Guide 12

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QIAGEN database workflow concept

STAR Q Punch AS: High-throughput punching and assay setup for QIAGEN STR Kits 13

A single, easily implemented workflow

• Up to 384 samples per day, 1 operator (96 samples/~90 minutes)

• Integrating card handling, imaging, punching and liquid handling into one workflow

• Identification of samples and calculation of optimal punch position

• Full traceability of samples including export to and import from LIMS systems

Investigator GO! PCR Kits 24plex GO! IDplex GO! ESSplex SE GO!

easyPunchfor punching and PCR setup

QIAgility for CE setupInvestigator STR GO! Punch Buffer

Sample to Insight

Automated EasiCollect/Bode Buccal card workflow


Validated automated punching and PCR setup with the easyPunch

• Fully automated punching of collection cards and reaction setup

• Validated setup protocols for Investigator 24plex GO! Kit

• ~90 min for 96 samples

True walk away automation for optimal results and processes using collection cards and Investigator 24plex GO! Kit

Sample to Insight

Features and Benefits

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Sample types compatible with the STAR Q Punch AS

Supplier ProductGE Healthcare EasiCollect (indicating FTA cards with cardboard frame, 5cm x 5cm)

Copan NUCLEIC-CARDs (indicating cards with cardboard frame, 51mm x 51mm)

Bode Bode Buccal 2

STAR Q Punch AS: High-throughput punching and assay setup for QIAGEN STR Kits

Sample to Insight

Collection card workflow

1616

Cards are inserted into carriers before

loading on the instrument.

Instrument performs visualization of the stained area of the

FTA card and punches out a 1.2mm section into a 96-well

plate.

Instrument performs assay set-up for STR

PCR using pre-defined protocols for QIAGEN direct amp

Investigator STR GO! kits.

Reagent tubes and pipette tips are

refilled between each run.

Basic principle

384 samples/day with a single analyst as part of an integrated workflow with QIAgility for downstream CE set-up.

Sample files are available for export after PCR set-up

STAR Q Punch AS: High-throughput punching and assay setup for QIAGEN STR Kits

Sample to Insight

Blood on FTA cards


Validation with Investigator 24plex GO! Kit

Study setup and results• 36 cards from different donors

◦ 5 replicates each, total of 180 samples

• 100 % full profiles at 200 RFU threshold

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Cross contamination study


Setup of contamination study:

No allelic peaks in negative samples demonstrated

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Automation of swabs


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Database swab workflow

The STAR Q Instruments for High Throughput automation of sample prep and assay setup 20

A single, easily implemented workflow

Investigator GO! PCR Kits• 24plex GO! Kit• IDplex GO! Kit• ESSplex SE GO! Kit

Hamilton STARletfor swab lysis and PCR setup

QIAgility for CE setup

• Up to 384 samples per day as part of a full workflow from sample to CE-setup, 1 operator

◦ 528 max instrument capacity/day (stand alone, 3 runs x 2 plates with total 176 samples/run)

• Integrating swab lysis and liquid handling in one workflow

• Full traceability of samples including export to and import from LIMS systems

Investigator STR GO! Lysis Buffer

Sample to Insight

Swab workflow

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Basic principle STAR Q Swab AS

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Buccal swabs manually loaded into Deep Well Plate and plate loaded

on to the instrument.

Instrument dispenses lysis buffer and

performs heating/shaking of

samples

Instrument performs assay set-up for STR PCR using predefined protocols for QIAGEN direct amp Investigator STR GO! kits. Aliquot of sample is added to

reaction plate

Reagent troughs and pipette tips are refilled

between each run.

Sample files are available for export after PCR set-up

The STAR Q Instruments for High Throughput automation of sample prep and assay setup

384 samples/day with a single analyst as part of an integrated workflow with QIAgility for downstream CE set-up.

Sample to Insight

Features and Benefits

The STAR Q Instruments for High Throughput automation of sample prep and assay setup

Sample types compatible with the STAR Q Swab AS

Manufacturer Name and description Reference Number

GE Healthcare Omni Swab WB100035

Copan Flocked Swab 552C

Sarstedt Cotton Swab 80.629

Puritan Polyester Swab 25-806

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Puritan Polyester swabGEHC Omni swab

Copan Flocked Swab

Sarstedt Cotton swab

Sample to Insight

Buccal cells on Omni swab

The STAR Q Instruments for High Throughput automation of sample prep and assay setup 23

Validation with Investigator 24plex GO! Kit

Study setup and results• Swabs from 20 different donors in 4 replicates each

• Checkerboards with alternating water samples

◦ Total of 80 buccal samples and x-contamination controls

• 100 % full profiles at 200 RFU threshold

• No sample carry over observed

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Literature and Resources


Application notes

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Summary


QIAGEN’s database workflow provides:High first pass success rates with 24plex

Unique Quality Sensor for informed rework strategy

Protocols and automation for any sample type

CE setup automation for increased throughput

Experienced forensic scientists to assist with validation

Sample to Insight

Thanks!


Investigator 24plex GO! · Blood on FTA cards STAR Q Punch AS: High -throughput punching and assay...

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Transcript of Investigator 24plex GO! · Blood on FTA cards STAR Q Punch AS: High -throughput punching and assay...