June 2022

QIAcard® FTA Elute Buffer

Handbook
For elution of nucleic acids from QIAcard FTA
Elute formats

Sample to Insight
Contents
Kit Contents ............................................................................................................... 3

Shipping and Storage ................................................................................................. 3

Intended Use .............................................................................................................. 3

Safety Information ....................................................................................................... 4

Quality Control........................................................................................................... 4

Introduction ................................................................................................................ 5

Equipment and Reagents to Be Supplied by User ............................................................ 6

Protocol: Procedure for QIAcard FTA Elute Formats Spotted with Extracted DNA ................ 7

Protocol: Procedure for QIAcard FTA Elute Formats Spotted with Biological Fluids ............ 12

Troubleshooting Guide .............................................................................................. 16

Ordering Information ................................................................................................ 17

Document Revision History ......................................................................................... 18

2 QIAcard FTA Elute Buffer Handbook 06/2022

Kit Contents
QIAcard FTA Elute Buffer

Catalog no. WB120100

Buffer volume 40 ml

QIAcard FTA Elute Buffer (40 ml) 1 bottle

Shipping and Storage

The QIAcard FTA Elute Buffer is shipped at room temperature (15–25°C) and should be stored
at 2–8°C upon arrival. Stored properly, the QIAcard FTA Elute Buffer is stable until the
expiration date indicated on the product.

Samples on QIAcard FTA Elute formats can be stored in a multi-barrier pouch with desiccant for
long term storage.

Intended Use
The QIAcard FTA Elute Buffer enables improved recovery of nucleic acids from QIAcard FTA
Elute formats. The buffer improves the elution efficiency of nucleic acids from various inputs,
including purified DNA, blood and saliva, applied to QIAcard Elute formats. This product is
not intended for the diagnosis, prevention or treatment of a disease.

Handling: Always wear gloves to avoid contamination of QIAcard FTA Elute formats. Follow
standard precautions when handling biological specimens.

QIAcard FTA Elute Buffer Handbook 06/2022 3

Safety Information
When working with chemicals, always wear a suitable lab coat, disposable gloves, and
protective goggles. For more information, please consult the appropriate safety data sheets
(SDSs). These are available online in convenient and compact PDF format at
www.qiagen.com/safety, where you can find, view and print the SDS for each QIAGEN® kit
and kit component.

Quality Control
In accordance with QIAGEN’s ISO-certified Quality Management System, each lot of the
QIAcard FTA Elute Buffer is tested against predetermined specifications to ensure consistent
product quality.

4 QIAcard FTA Elute Buffer Handbook 06/2022

Introduction
Flinders Technology Associates (FTA) is a chemically coated cellulose matrix that is capable
of lysing cells upon contact and denaturing proteins.

Normal QIAcard FTA formats are not intended for elution of nucleic acids and are regularly
used during a direct PCR analysis process. If nucleic acids need to be released from FTA cards,
we recommend using QIAcard FTA Elute formats:

• QIAcard FTA Elute Indicating Micro (cat. no. WB120411 or WB120412) for clear
biological fluids or purified DNA

• QIAcard FTA Elute Micro (cat. no. WB120401 or WB120410) for colored
biological fluids

QIAcard FTA Elute formats are designed for room temperature (15–25°C) shipment,
preservation and subsequent elution of previously released nucleic acids from biological
samples. Sample material, such as cells, bacteria and blood, are lysed upon contact with the
cards. By using the QIAcard FTA Elute Buffer, nucleic acids can be eluted from the card.

Extracted DNA or several other sample materials are routinely stored frozen for sample
archiving. QIAcard FTA Elute can be used to stabilize and protect nucleic acids for long-term
storage at room temperature. This procedure describes how to apply, store and elute nucleic
acids on the above-described cards using QIAcard FTA Elute Buffer.

QIAcard FTA Elute Buffer Handbook 06/2022 5

Equipment and Reagents to Be Supplied by User
When working with chemicals, always wear a suitable lab coat, disposable gloves and
protective goggles. For more information, consult the appropriate safety data sheets (SDSs),
available from the product supplier.

 QIAcard FTA Elute buffer

 TE−4 buffer (10 mM Tris·Cl, 0.1 mM EDTA)
 Recommended: QIAshredder (cat. no. 79656)
 UniCore Punches 3.00 mm (cat. no. WB100078 or WB100039)
 Optional: UniCore Punches 6.00 mm (cat. no. WB100082 or WB100040)
 Cutting Mat 6.0" x 8.0" or 2.5" x 3.0" (cat. no. WB100020 or WB100088)
 Heated mixer or shaker
 2 ml microcentrifuge tubes

Additional material required for “Procedure for QIAcard FTA Elute formats spotted
with biological fluids”

 QIAGEN Proteinase K (cat. no. 19131)

6 QIAcard FTA Elute Buffer Handbook 06/2022

Protocol: Procedure for QIAcard FTA Elute
Formats Spotted with Extracted DNA
This protocol is for elution of DNA from QIAcard FTA Elute cards that are spotted with
previously extracted DNA using QIAcard FTA Elute Buffer. For clear biological fluids or
extracted DNA, we recommend using QIAcard FTA Elute Indicating Micro (cat. no.
WB120411 or WB120412) as the included purple dye turns white when liquid sample
material is applied. The color change enables an efficient sample visualization for subsequent
sample punching.

Applying DNA extract to QIAcard FTA Elute Indicating Micro

1. Label the QIAcard FTA Elute Indicating Micro with the appropriate sample identification.
2. Pipette a volume of up to 75 µl DNA extract onto the card within the printed circle.
3. Allow the sample to air dry at least 3 h at room temperature until dry.
4. Samples applied to QIAcard FTA Elute Indicating Micro should be archived at room
temperature (15–25°C) in a Multi-Barrier Pouch (cat. no. WB100036) with an Indicating
Desiccant Pack (cat. no. WB100003) or stored in a humidity-controlled, cool, dry
environment.

QIAcard FTA Elute Buffer Handbook 06/2022 7

Elution of DNA from QIAcard FTA Elute Indicating Micro

1. Place the QIAcard FTA Elute Indicating Micro on a cutting mat.

2. Remove up to four 3 mm punches from the FTA Elute Card, and place the punches into a
single 2 ml microcentrifuge tube. To prevent contamination, we recommend using a
clean punch (remove and discard a punch from an unused QIAcard FTA Elute card)
between card-to-card sample punching.
Note: One to four 3 mm punches can be used based on known sample
concentration and DNA input required for amplification. A volume of 25 µl spotted
DNA is sufficient for four 3 mm punches. As an alternative, a volume of 75 µl
spotted DNA is sufficient for seven 6 mm punches.
Note: Representative examples for 25 µl and 75 µl fluid spotted on QIAcard FTA Elute
cards are shown in Figure 1.
Optional: If 3 mm punches are used, the punches can alternatively be washed within a
QIAshredder basket. For 6 mm punches, up to four punches can be handled this way.
Place the punches into QIAshredder baskets.
IMPORTANT: Before continuing with the next step, read the instruction below carefully:
If the punches are left in TE−4 buffer too long, the sample can potentially be
compromised. Therefore, when processing more than two samples, we recommend
processing only two samples at a time for step 3–6: process only the first two samples,
and then process the next batch of two samples and continue processing samples in sets
of two. Once the first wash is completed for all samples, proceed with the next wash
(step 7) in sets of two samples until all samples are ready for step 8.
3. Pipette 500 µl TE−4 buffer into the microcentrifuge tube containing the punches.
Optional: If the punches are washed within a QIAshredder basket, pipette 500 µl
TE−4 buffer into the QIAshredder basket containing the punches.

8 QIAcard FTA Elute Buffer Handbook 06/2022

 4. Close the microcentrifuge tube and vortex for 5 s. Ensure the punches move up into the
center of the microcentrifuge tube when they are vortexed.
Optional: If the punches are washed within a QIAshredder basket, close the basket
and ensure the punches move up into the center of the basket when they are vortexed.
5. Briefly centrifuge the microcentrifuge tubes to remove any excess liquid from the cap.
Optional: If the punches are washed within a QIAshredder basket, centrifuge at
2000 rpm for 1 min to separate the wash liquid from the punches.
6. Remove excess TE−4 buffer and discard.
Optional: If the punches are washed within a QIAshredder basket, remove the TE–4 buffer
flowthrough from the microcentrifuge tube.
7. Repeat steps 3–6 once or twice (for a total of two to three washes with TE−4 buffer).
Note: If high concentration of inhibitors is expected, we recommend washing three times.
Optional: If the punches were washed within a QIAshredder basket, transfer the
punches into a fresh 2 ml microcentrifuge tube. Retain the QIAshredder basket inside
the original microcentrifuge tube.
8. Pipette an appropriate amount of QIAcard FTA Elute Buffer into the microcentrifuge tube
containing the sample punches based on the number of punches and suggested volumes
in Table 1.
Note: The QIAcard FTA Elute Buffer serves as the elution buffer.
Note: Seven 6 mm punches require a volume of 400 µl QIAcard FTA Elute Buffer.
9. Place the microcentrifuge tube on a heated mixer or shaker at 95°C for 20 min at
1000 rpm.
10. After incubation, briefly centrifuge the microcentrifuge tube to remove any excess liquid
from the cap.

QIAcard FTA Elute Buffer Handbook 06/2022 9

11. Place a clean spin basket into a new microcentrifuge tube. Transfer the punches and
eluate to the spin basket and spin at maximum speed for 2 min. Remove the spin basket,
discard the punches, and proceed with quantification and/or amplification.
Recommended: To obtain the best results, we recommend transferring the punches and
the eluate to a QIAshredder basket. Place the basket back into the tube that was used for
elution on the heated mixer or shaker. Close the basket and spin at maximum speed for
2 min. Remove the QIAshredder basket, discard the punches, and proceed with
quantification and/or amplification.
Optional: If the punches were washed within a QIAshredder basket, the same
QIAshredder basket can be used for step 11.
Note: If there is still liquid in the basket, use a clean pipette tip to move the punches
away from the filter of the QIAshredder basket and repeat centrifugation.
Note: Concentration using a suitable method, such as using filtration devices, may be
required for any combination of punches and elution volumes, depending on the initial
amount of the sample.. Concentrating the eluate by evaporation is strongly discouraged.
12. Store extracts according to your laboratory protocols.
Table 1. Recommended elution volume (QIAcard FTA Elute Buffer) for different numbers of 3 mm or 6mm card punches.

QIAcard FTA Elute Buffer QIAcard FTA Elute Buffer

Number of punches for Ø 3 mm punches for Ø 6 mm punches

1 ≥50 µl ≥100 µl

2 ≥75 µl ≥160 µl

3 ≥100 µl ≥200 µl

4 ≥125 µl ≥260 µl

5 ≥150 µl ≥320 µl

6 ≥175 µl ≥360 µl

7 ≥200 µl ≥400 µl

Concentration may be required for any combination of punches and elution volumes dependent

10 QIAcard FTA Elute Buffer Handbook 06/2022

Figure 1. Representative examples for 25 µl and 75 µl DNA spotted on QIAcard FTA Elute cards. When applying 25 µl,
up 4 x 3 mm punches can be removed from the application area of the card. When applying 75 µl, up 7 x 6 mm
punches can be removed from the application area of the card.

QIAcard FTA Elute Buffer Handbook 06/2022 11

Protocol: Procedure for QIAcard FTA Elute
Formats Spotted with Biological Fluids
This protocol is for elution of nucleic acids from QIAcard FTA Elute cards that are spotted with
biological fluids (e.g., blood, saliva) using QIAcard FTA Elute Buffer.

For clear biological fluids, such as saliva, we recommend using QIAcard FTA Elute Indicating
Micro (cat. no. WB120411 or WB120412) as the included pink dye turns white when liquid
sample material is applied. The color change enables an efficient sample recognition for
subsequent sample punching.

For colored biological fluids, such as blood, we recommend using the non-indicating QIAcard
FTA Elute Micro (cat. no. WB120401 or WB120410).

Applying biological fluids to QIAcard FTA Elute cards

1. Label the QIAcard FTA Elute format with the appropriate sample identification.
2. Pipette a volume of up to 75 µl biological fluids onto the card within the printed circle.
3. Allow the sample to air dry at least 3 h at room temperature until dry.
Samples applied to QIAcard FTA Elute Micro formats should be archived at room
temperature (15–25°C) in a Multi-Barrier Pouch (cat. no. WB100036) with an Indicating
Desiccant Pack (cat. no. WB100003) or stored in a humidity-controlled, cool, dry
environment.

12 QIAcard FTA Elute Buffer Handbook 06/2022

Elution of nucleic acids derived from biological fluids from QIAcard FTA Elute
formats

1. Place the QIAcard FTA Elute card on a cutting mat.

2. Remove the desired number of punches (see Table 2) from the FTA Elute Card and place
the punches into a single 2 ml microcentrifuge tube. To prevent contamination, we
recommend using a clean punch (remove and discard a punch from an unused
QIAcard FTA Elute card) between card-to-card sample punching.
Note: Representative examples for 25 µl and 75 µl fluid spotted on QIAcard FTA
Elute cards are shown in Figure 1.
Optional: If 3 mm punches are used, the punches can alternatively be washed within
a QIAshredder basket. For 6 mm punches, up to four punches can be handled this
way. Place the punches into QIAshredder baskets.
IMPORTANT: Before continuing with the next step, read the instruction below carefully:
If the punches are left in TE−4 buffer too long, the sample can potentially be
compromised. Therefore, when processing more than two samples, we recommend
processing only two samples at a time for step 3–6: process only the first two samples,
and then process the next batch of two samples and continue processing samples in sets
of two. Once the first wash is completed for all samples, proceed with the next wash
(step 7) in sets of two samples until all samples are ready for step 8.
3. Pipette 500 µl TE−4 buffer into the microcentrifuge tube containing the punches.
Optional: If the punches are washed within a QIAshredder basket, pipette 500 µl
TE−4 buffer into the QIAshredder basket containing the punches.
4. Close the microcentrifuge tube and vortex for 5 s. Ensure the punches move up into the
center of the microcentrifuge tube when they are vortexed.
Optionally: If the punches are washed within a QIAshredder basket, close the basket
and ensure the punches move up into the center of the basket when they are vortexed.

QIAcard FTA Elute Buffer Handbook 06/2022 13

 5. Briefly centrifuge the microcentrifuge tubes to remove any excess liquid from the cap.
Optional: If the punches are washed within a QIAshredder basket, centrifuge at
2000 rpm for 1 min to separate the wash liquid from the punches.
6. Remove excess TE−4 buffer and discard.
Optional: If the punches are washed within a QIAshredder basket, remove the TE–4
buffer flowthrough from the microcentrifuge tube.
7. Repeat steps 3–6 once or twice (for a total of two to three washes with TE−4 buffer).
Note: If high amount of inhibitors are expected, we recommend washing three times.
Optional: If the punches were washed within a QIAshredder basket, transfer the
punches into a fresh 2 ml microcentrifuge tube. Retain the QIAshredder basket inside
the original microcentrifuge tube,
8. Pipette an appropriate amount of QIAcard FTA Elute Buffer and Proteinase K into the
microcentrifuge tube containing the sample punches based on the number of punches
and suggested volumes in Table 2
Note: The QIAcard FTA Elute Buffer and Proteinase K serves as the elution buffer.
Note: Seven 6 mm punches require a volume of 400 µl QIAcard FTA Elute Buffer.
9. Place the microcentrifuge tube on a heated mixer or shaker at 60°C (Proteinase K
digestion) for 25 min at 1000 rpm.
10. For inactivation of Proteinase K, place the microcentrifuge tube on a heated mixer or
shaker at 95°C (Proteinase K inactivation) for 5 min at 1000 rpm.
11. After incubation, briefly centrifuge the microcentrifuge tube to remove any excess liquid
from the cap.

14 QIAcard FTA Elute Buffer Handbook 06/2022

12. Place a clean spin basket into a new microcentrifuge tube. Transfer the punches and
eluate to the spin basket and spin at maximum speed for 2 min. Remove the spin basket,
discard the punches, and proceed with quantification and/or amplification.
Recommended: To obtain the best results, we recommend transferring the punches to a
QIAshredder basket. Place the basket back into the tube that was used for elution on the
heated mixer or shaker. Pipette the eluate onto the sample punches, close the basket and
spin at maximum speed for 2 min. Remove the QIAshredder basket, discard the punches,
and proceed with quantification and/or amplification.
Optional: If the punches were washed within a QIAshredder basket, the same
QIAshredder can be used for elution.
Note: If there is still liquid in the basket, use a clean pipette tip to move the punches
away from the filter of the QIAshredder basket and repeat centrifugation.
Note: Concentration using a suitable method, such as using filtration devices, may be
required for any combination of punches and elution volumes, depending on the initial
amount of the sample. . Concentrating the eluate by evaporation is strongly discouraged.
13 . Store extracts according to your laboratory protocols.
Table 2. Recommended elution volume (QIAcard FTA Elute Buffer) for different numbers of 3 mm and 6 mm card
punches.

QIAcard FTA Elute

Number of Proteinase K Buffer Proteinase K QIAcard FTA Elute Buffer
punches for Ø 3 mm punches for Ø 3 mm punches for Ø 6 mm punches for Ø 6 mm punches

1 4 µl ≥50 µl 6 µl ≥100 µl

2 5 µl ≥75 µl 8 µl ≥160 µl

3 6 µl ≥100 µl 10 µl ≥200 µl

4 7 µl ≥125 µl 14 µl ≥260 µl

5 8 µl ≥150 µl 16 µl ≥320 µl

6 9 µl ≥175 µl 18 µl ≥360 µl

7 10 µl ≥200 µl 20 µl ≥400 µl

Concentration may be required for any

QIAcard FTA Elute Buffer Handbook 06/2022 15

Troubleshooting Guide
This troubleshooting guide may be helpful in solving any problems that may arise. For more
information, see also the Frequently Asked Questions page in our Technical Support Center:
www.qiagen.com/FAQ/FAQList.aspx. The scientists in QIAGEN Technical Services are
always happy to answer any questions you may have about either the information or protocols
in this handbook (for contact information, visit support.qiagen.com).

Comments and suggestions

Calculating the percent 1. Measure the diameter of the sample area (stain) covered by the DNA extract and
recovery of eluted DNA divide this value by 2 to calculate the radius (mm).
2. Calculate the area of the stain as follows:
Area of stain (mm2) = 3.14 × radius of stain (mm) × radius of stain (mm)
For a sample area with a 10 mm diameter:
Area of 10 mm stain (mm2) = 3.14 × 5 mm × 5 mm = 78.5 mm2
3. Calculate quantity of DNA per mm2 of stain as follows:
Amount of DNA spotted onto FTA Elute card (pg)/area of stain (mm2)
For a 25 µl sample at a concentration of 1,000 pg/µl:
25,000 pg DNA applied per 78.5 mm2 = 25,000 pg /78.5 mm2
Quantity of DNA per mm2 of stain = 318.47 pg DNA/mm2
4. Calculate the area of a 3 mm punch.
Area of 3 mm punch (mm2) = 3.14 x radius of punch (mm) x radius of punch (mm)
Area of 3 mm punch (mm2) = 3.14 x 1.5 mm x 1.5 mm
Area of one, 3 mm punch = 7.065 mm2
Area of two 3 mm punches = 2 x 7.065 mm2 = 14.13 mm2
5. Calculate quantity of DNA per 3 mm punch:
If DNA was applied to the FTA Elute card at 318.47 pg/mm2 and the area of each
3 mm punch is 7.065 mm2, each 3 mm punch contains:
318.47 pg DNA/mm2 × 7.065 mm2
Quantity of DNA per 3 mm punch = 2,250 pg
(Multiply 2,250 pg by number of 3 mm punches processed)
6. Calculate % DNA recovery:
% DNA recovery = 100 × (A)/(B)
Where:
A = quantity of DNA eluted from FTA Elute Card (total pg)
B = expected quantity of DNA per 3 mm punch, calculated in step 5 (pg)
Use of QIAGEN Lyse & We do not recommend using the Investigator Lyse & Spin Basket, as the separation
Spin Baskets performance of the recommended QIAshredder baskets is superior for this application.
Use of Investigator Lyse When using Investigator Lyse & Spin Baskets, the punched discs may seal the holes of the
& Spin Baskets for basket during centrifugation and prevent lysate flowthrough. However if the use of
separation not working Investigator Lyse & Spin Basket is required in your laboratory, it is advised to push the
as expected punched discs towards the wall of the basket. We recommend using the QIAshredder
baskets for separation instead.

16 QIAcard FTA Elute Buffer Handbook 06/2022

Ordering Information

Product Description Cat. no.

QIAcard FTA Elute Buffer (40 ml) 40 ml QIAcard FTA Elute WB120100
Buffer.

QIAcard FTA Elute Indicating Micro 100 QIAcard FTA Elute WB120411
(25) Indicating cards (one sample
area per card).

QIAcard FTA Elute Indicating Micro 25 QIAcard FTA Elute WB120412

(25) Indicating Micro cards (one
sample area per card).

QIAcard FTA Elute Micro (25) 100 QIAcard FTA Elute WB120410
Micro cards (four sample
areas per card).

QIAcard FTA Elute Micro (25) 25 QIAcard FTA Elute Micro WB120401
cards (4 sample areas per
card).

UniCore Punch Kit 3.0 mm (4) 4 UniCore Punches, 3.0 mm, WB100039
and 2 cutting mats.

UniCore Punch 3.0 mm (25) 25 UniCore Punches, 3.0 WB100078

mm.

UniCore Punch Kit 6.00 mm (4) 4 UniCore Punches, 6.0 mm, WB100040
and 2 cutting mats.

UniCore Punches 6.00 mm (25) 25 UniCore Punches, 6.0 WB100082

mm.

Cutting Mat 2.5" × 3.0" 1 Cutting Mat 2.5" × 3.0". WB100088

Cutting Mat 6.0" × 8.0" 1 Cutting Mat, 6.0" × 8.0". WB100020

QIAcard FTA Elute Buffer Handbook 06/2022 17

Product Description Cat. no.

Multi-Barrier Pouches, 3.75” x 3” 100 Multi-Barrier Pouches, WB100036

(100) 3.75” x 3”.

Indicating Desiccant Pack (1000) 1000 Indicating Desiccant WB100003

Packs (1 g each)

QIAshredder (250) 250 disposable cell-lysate 79656

homogenizers for use in
nucleic acid minipreps, caps

QIAGEN Proteinase K (2 ml) 2 ml (>600 mAU/ml, 19131

solution)

For up-to-date licensing information and product-specific disclaimers, see the respective
QIAGEN kit handbook or user manual. QIAGEN kit handbooks and user manuals are
available at www.qiagen.com or can be requested from QIAGEN Technical Services or your
local distributor.

Document Revision History

Date Changes

06/2022 Initial release

18 QIAcard FTA Elute Buffer Handbook 06/2022

Limited License Agreement

Use of this product signifies the agreement of any purchaser or user of the product to the following terms:
1. The product may be used solely in accordance with the protocols provided with the product and this handbook. QIAGEN grants no license under any of its
intellectual property to use or incorporate the product except as described in the protocols provided with the product, this handbook, and additional protocols
available at www.qiagen.com. Some of these additional protocols have been provided by QIAGEN users for QIAGEN users. These protocols have not been
thoroughly tested or optimized by QIAGEN. QIAGEN neither guarantees them nor warrants that they do not infringe the rights of third-parties.
2. Other than expressly stated licenses, QIAGEN makes no warranty that this product and/or its use(s) do not infringe the rights of third-parties.
3. This product are licensed for one-time use and may not be reused, refurbished or resold.
4. QIAGEN specifically disclaims any other licenses, expressed or implied other than those expressly stated.
5. The purchaser and user of the product agree not to take or permit anyone else to take any steps that could lead to or facilitate any acts prohibited above.
QIAGEN may enforce the prohibitions of this Limited License Agreement in any Court, and shall recover all its investigative and Court costs, including attorney
fees, in any action to enforce this Limited License Agreement or any of its intellectual property rights relating to the product and/or its components.
For updated license terms, see www.qiagen.com.
06/2022 HB-3091-001 © 2022 QIAGEN, all rights reserved.

Trademarks: QIAGEN®, Sample to Insight®, QIAcard® (QIAGEN Group).

QIAcard FTA Elute Buffer Handbook 06/2022 19

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