HBV PCR

AccuPower®
HBV Quantitative PCR Kit
HBV-1111
Quantitative test kit

Hepatitis B virus DNA
AccuPower® HBV Quantitative PCR Kit
User’s Guide
Version No.: 2.2 (2012-12)
BIONEER CORPORATION
8-11, Munpyeongseo-ro, Daedeok-gu,
Daejeon, 306-220, Republic of Korea
Tel: +82-1588-9788
Fax: +82-42-930-8600
Email: order@bioneer.co.kr
www.bioneer.com
1
Safety Warnings and Precautions
Please inquire BIONEER’s Customer Service Center to obtain a copy of

the Material Safety Data Sheet (MSDS) for this product.
Before, during and after use of this kit as described in this User’s
Guide, all potentially hazardous materials (i.e. materials that may have
come in contact with clinical samples) including tubes, tips and
materials should be processed and disposed of according to
applicable and appropriate regulations of the municipality/
government in which this product is being used.
Please read the User’s Guide before using this Kit. Please check the
integrity of all tubes, tips and other materials supplied with this kit
prior to use. Adhere to general clinical laboratory safety procedures
during the experiment.
Some applications that may be performed with this kit may infringe
upon existing patents in certain countries. The purchase of this kit
does not include or provide a license to perform patented
applications. Users may be required to obtain a license depending on
country and application. We do not condone nor recommend the
unlicensed use of a patented application.
Warranty and Liability
All BIONEER products are manufactured and tested under strict

quality control protocols. BIONEER guarantees the quality of all
directly manufactured products during the warranty period of one (1)
year from date of purchase. If any issues are discovered relating to
compromise in product quality, immediately contact BIONEER’s
Customer Service Center (order@bioneer.com).
BIONEER does not assume liability for misuse of the product, i.e.
usage of the product for any purposes other than its intended
purpose as described in the appropriate and applicable User’s Guide.
2
BIONEER assumes liability under the condition that the user discloses
all information related to the problem to BIONEER in written form
within 30 days of occurrence.
Trademarks
AccuPower® is a registered trademark of BIONEER Corporation,

Republic of Korea. Exicycler™, ExiSpin™ and ExiPrep™ are
trademarks of BIONEER Corporation, Republic of Korea.
FAM and TAMRA are trademarks of Applera Corporation.
Excel™ is a trademark of Microsoft Corporation.
Copyright © 2012 by Bioneer Corporation. All Rights Reserved.
3
TABLE OF CONTENTS
1. INTENDED USE……………………………………………..…………....6
2. INTRODUCTION……………………..…………………………..…….7
3. FEATURES AND PRINCIPLE OF THE TEST……….………….8
3.1. Kit Format………….………...……………………..…..........……....9
3.2. Sensitivity and Dynamic Range..………...………………..…...9
3.3. Internal Positive Control (IPC)……….………………………...9
4. CONTENTS OF THE KIT……………………………….……………..10
5. STORAGE………………….……………….……………………………..11
6. REQUIRED MATERIALS AND EQUIPMENT…………………....12
7. GENERAL PRECAUTIONS.……………………………………13
8. PROTOCOL….…………….………………………………………..…..14
8.1. Preparation: Specimen Collection, Storage, and
Transport…………………………………….………………………14
8.1.1. Specimen Collection.…………….….…………………….14
8.1.2. Sample Storage..…..……………………..……………………..14
8.1.3. Sample Transport………………………………………….15
8.1.4. Interfering Substances…………………………………….15
8.2. Work Flow Schema………..……………………………………..16
8.2.1. Nucleic Acid Extraction………..……...……………………...17
8.2.2. PCR Preparation………………………..…………..………..18
4
8.2.3. P r o g r a m m i n g t h e E x i c y c l e r ™ 9 6 R e a l - T i m e
Quantitative Thermal Block……………………...………….21
9. DATA ANALYSIS.….…….……………………………………………..27
9.1. Process of Data Analysis...……………………………...……..27
9.2. Analysis Results…………………………………...……….…31
10. TROUBLESHOOTING…………...…………………………………….33
11. REFERENCES …………………………………………………………..35
12. SYMBOLS…………………………………………………………….…...36
5
1. INTENDED USE
The AccuPower® HBV Quantitative PCR Kit is a ready-to-use,

all-in-one-tube kit designed for the quantification of hepatitis B
virus (HBV) DNA within clinical samples through qPCR.
This kit is optimized for use with BIONEER’s Exicycler™ 96

Real-Time Quantitative Thermal Block (Cat. No: A-2060).
The use of kit is only for qualified users trained to correctly

and safely handle clinical specimens and conduct molecular
biological experiments. All waste generated before, during and
after the experiment should be processed in compliance with
local regulations.
Please read the instructions contained within this manual

thoroughly before using this Kit and check the integrity of all
components before use.
6
2. INTRODUCTION
Hepatitis B Virus (HBV) is reported to be closely related to

acute/chronic hepatitis, liver cirrhosis and liver cancer. Most
healthy adults are able to rid the virus through a natural
immune reaction. But individuals such as children with weak
immune systems or newborns that have been infected by their
mothers have been reported to develop hepatitis B with 90%
incidence.
HBV infection can be determined by detecting HBsAg (HBV

surface antigen) in serum. But in order to distinguish between
chronic and active infection, one must quantify circulating HBV
DNA levels and liver enzyme levels.
AccuPower® HBV Quantitative PCR Kit is designed to quantify

HBV virus DNA extracted from clinical blood samples.
Quantification through real-time PCR has several advantages
over conventional PCR including superior sensitivity, specificity
and one less step of post-PCR processing. Also, because the
assay is quantitative, this allows physicians to validate
treatment efficacy and offer more effective treatment.
7
3. FEATURES AND PRINCIPLE OF THE TEST

Real-time PCR involves the selective amplification of a target
sequence while monitoring the progress of amplification in real-
time through a visualizing agent, such as a fluorescent dye. The
specificity is provided by a pair of specific primers, along with a
hydrolysis probe which is also sequence specific. Monitoring
amplified product is conducted by labeling the hydrolysis probe
with a matched pair of fluorescent dyes (5’-FAM; 3’- QUENCHER).
Due to fluorescence resonance energy transfer (FRET), an intact
probe will not emit light. However, upon cleavage by the 5’ – 3’
exonuclease activity of the DNA polymerase, during PCR, FAM
will emit a specific wavelength of light within the visible
spectrum (520 nm) when cleaved after binding to the amplicon.
The Kit was designed to maximize reproducibility and ease-of-
use by vacuum-drying the primers, probes, DNA polymerase,
dNTPs and salts using our proprietary stabilization technology to
preserve the full activity of the mixed reagents. The primer-
probe set was selected from a pool of primer-probe
combinations designed by our bioinformatics algorithms, to 1)
achieve maximum amplification efficiency, and 2) as with all of
our other AccuPower® Diagnostic Kits, match the thermal
cycler program so that this Kit could be run simultaneously with
other Kits from our AccuPower® Diagnostic Kit series.
8
3.1. Kit Format
AccuPower® HBV Quantitative PCR Kit is a ready-to-use

product that can analyze up to 96 samples (including controls)
in a single run.
3.2. Sensitivity and Dynamic Range
The sensitivity and the dynamic range of AccuPower® HBV

Quantitative PCR Kit are 9.55 IU/㎖ and 50 ~ 1X108 IU/㎖,
respectively.
3.3 Internal Positive Control (IPC)
AccuPower® HBV Quantitative PCR Kit employs Internal

Positive Control (IPC) amplification in all wells to confirm
correct PCR amplification.
9
4. CONTENTS OF THE KIT
Cat. No.
Labeling & Content
HBV-1111
HBV Premix 8 well-strip 12 x 8-well strips

①
in aluminum foil bag (96 tubes)
HBV Standard Positive Control 15 ul / tube
(2 x 102~106 copies/ul) x 8 strips
15 ul / tube
IPC DNA 2 x 8-well strips
②
(16 tubes)
15 ul / tube
DEPC DW (for NTC) 2 x 8-well strips
(16 tubes)
1800 ul / tube
③ DEPC DW
x 4 tubes
Optical sealing film 1 sheet
10
5. STORAGE
AccuPower® HBV Quantitative PCR Kit should

be stored at -22 ~ -18℃ away from
UV/sunlight. The Kit is guaranteed stable until
the expiration date printed on the label. Repeated freeze/thaw
cycles (more than once) should be avoided, as it may affect Kit
quality. If intermittent use of the kit is expected, only those
components that will be used should be removed from the
freezer. Reagents should not be stored at 4℃.
11
6. REQUIRED MATERIALS AND EQUIPMENT

(NOT PROVIDED IN THE KIT)
- Disposable powder-free gloves

- Appropriate volume Pipette set
- Sterilized pipette tips with filters
- 1.5 ml micro tubes or 15 ml conical tubes
- Equivalent protocol and materials necessary to extract DNA
(see 8.2.1. Nucleic Acid Extraction):
• AccuPrep® Viral DNA Extraction Kit (Cat. No.: K-3511,
Bioneer Corp., KOREA)
• ExiPrep™ Dx Viral DNA Kit (Cat. No.: K-4472, Bioneer
Corp., KOREA)
• ExiPrep™16 Dx (Cat. No.: A-5050, Bioneer Corp., KOREA)
- Vortex and microcentrifuge capable of handling 8-well

strip-tubes (e.g. ExiSpin™ Programmable Vortex/
Microcentrifuge (Cat. No: A-7040))
- Exicycler™ 96 Real-Time Quantitative Thermal Block
(Cat. No: A-2060, Bioneer Corp., Korea)
12
7. GENERAL PRECAUTIONS
- Store all kit components at -22 ~ -18℃.

- This kit is designed for the viral load
assay to quantify HBV viral DNA from blood and blood
products, not for the diagnosis of HBV infection.
- Always wear gloves and a mask when handling
biohazardous agents.
- DO NOT repeatedly freeze/thaw Kit components.
- DO NOT reuse opened reagents, nor mix reagents from
different production lots.
- DO NOT change the protocol as described in this manual.
- Always use sterile, filtered pipette tips.
- Clinical samples and their derivatives should be stored in a
separate location/freezer from where the rest of the Kit
components are stored.
- All Kit components should be allowed to slowly thaw for at
least 10 minutes before initiating an experiment.
- Briefly vortex and spin-down all Kit components after
thawing to ensure optimum results.
- All Stanadards should be added in a physically separate
location from where the premix is reconstituted.
13
8. PROTOCOL
8.1. Preparation: Specimen Collection, Storage, and

Transport
Caution: All samples should be treated as potential

biohazards.
Attention: For the best results, we recommend that DNA
extracts be derived from serum or plasma.
8.1.1. Specimen Collection
AccuPower® HBV Quantitative PCR Kit is optimized for DNA

extracted from serum or plasma sample. For serum or plasma
collection, standard specimen collection tubes such as BD SST
Serum Separator tubes or disposable tubes containing EDTA as
anticoagulant can be used.
Attention: All samples should be kept in preservative-free
containers.
8.1.2. Sample Storage
Do not store samples at 2~8℃ for over 7 day. For long term
storage of clinical samples, please aliquot and store at -20 ~ -
14
80℃.
8.1.3. Sample Transport
All samples should be transported in a shatterproof

transport container to prevent potential infection from sample
leakage. Samples should be transported according to
local/national guidelines regarding biohazard transportation.
8.1.4. Interfering Substances
Heparin (≥ 10 IU/㎖) is a known inhibitor of PCR. Samples

that have been collected in tubes containing heparin should
not be used. In addition, samples from heparin-treated
patients should not be used. Clinical samples may contain a
variety of PCR inhibitors. For efficient PCR, such inhibitors must
be removed during the DNA extraction and purification
process. Using Bioneer’s ExiPrep™ Dx Viral DNA Kit, will ensure
removal of PCR inhibitors and is recommended.
15
8.2. Work Flow Schema
16
8.2.1. Nucleic Acid Extraction
HBV DNA should be purified from raw clinical samples

before this real-time PCR experiment can be conducted. Please
perform viral DNA extraction according to pre-established
protocols, or, in the case of kit extraction of viral DNA, adhere
to the manufacturer protocol for DNA extraction.
* According to the extraction protocol or kit, real-time PCR

results can be varied. For the best results, we recommend the
following kits:
Products Cat. No. Platform
AccuPrep® Viral DNA Extraction Kit K-3511 Manual
ExiPrepTM Dx Viral DNA Kit K-4472 ExiPrepTM 16 Dx
17
8.2.2. PCR Preparation
① The appropriate number of premix tubes, controls and

nucleic acid extracts (if frozen) should be thawed for at least
10 minutes at room temperature. At least 6 tubes are needed
for experimental control. Therefore, the number of premix
tubes required for a single experiment is:
6 (1 NTC + 5 standards) + number of samples
② Briefly vortex and spin-down all reagents, controls and

samples prior to use.
③ Make a premix reconstitution mixture by combining 44 μl of

DEPC DW and 1 μl of Internal Positive Control (IPC) per
reaction. Make the mixture for at least one more tube than is
actually needed to account for tube adhesion loss of the
mixture. The following chart is a summary of reagent
requirements with 8 and 16 samples:
Number of samples 1 8 16
Non-Template Control
sample Standard positive control 5 ul Each 5ul Each 5ul
or clinical sample
Internal Positive Control 1 ul 9 ul 18 ul
DEPC DW 44 ul 396 ul 792 ul
Total Volume 50 ul 450 ul 900 ul
Volume per reaction 50 ul Each 50 ul Each 50 ul
18
④ Add 45 μl of the premix reconstitution mixture to all premix

tubes.
⑤ For the tubes that will be placed in A1 add 5 μl of DEPC DW

(purple tubes) as NTC. Please seal these wells with the
supplied optical adhesive cover to prevent contamination
from ensuing steps.
⑥ Move the tubes to a physically separated location before

proceeding. For the tubes that will be placed in B1~F1, add
5 μl of Standard positive control (natural strip).
⑦ Add 5 μl of nucleic acid extracts to the remaining tubes,

making note of which samples correspond to which original
clinical sample.
⑧ Cut the supplied Optical Film to size with scissors or a razor

and use the Optical Film Applicator to completely seal the
tubes. (Remember to keep track of the clinical samples)
⑨ To mix the tube thoroughly (to dissolve the Premix pellet and
mix the DNA), use the ExiSpin™ (Cat.No : A-7040, Bioneer
Corp., Korea) and perform 20 cycles at 2500 rpm for 5 sec.
19
spindown / 20 sec. hard vortex.
⑩ While the ExiSpin™ is operating, you can start the

Exicycler™ 96. (Refer to chapter 8.2.3. Programming the
Exicycler™ 96 Real-Time Quantitative Thermal Block)
⑪ After the ExiSpin™ cycling is complete, we recommend

placing the tubes in the Exicycler™ 96 immediately.
20
8.2.3. Programming the Exicycler™ 96 Real-Time

Quantitative Thermal Block
① Turn the Standby Power Switch, located at the rear of the

instrument ON. The front ring-LED status light should turn
on RED (Fig.1-①).
Fig. 1) Front view of the instrument showing operating buttons
② Press the front Operation Power Switch for 2 seconds. A brief

self test sequence will initiate. If the self test passes, the front
ring-LED will blink GREEN (Fig.1-②), with a short beep.
21
③ Push the Door Switch for 2 seconds to slide the 96-well

thermal block out. Insert the reaction tubes in their pre-
determined locations. (Remember, the NTC tube go in well
A1 and the standards tubes go in wells B1~F1) After sample
loading is complete, push the Door Switch for 2 seconds to
close the door.
Fig. 2) Side view and the 96-well thermal block of the instrument
④ Use a spreadsheet program capable of creating ‘*.xls’ files

and create a sample layout map (Fig. 3).
Fig. 3) Sample Layout Map
22
⑤ Save the sample layout map file as an ‘*.xls’ file. Save the file
in an easily findable location.
⑥ Double click the ‘ExiDiagnosis Run’ icon to start the

program. If a ‘Device is Not ready’ warning appears, please
close and restart the program. If a ‘System is ready’ prompt
appears, the computer has successfully connected to the
instrument.
⑦ To setup the protocol for PCR, click ‘File - Design

Experiment’ from the top menu bar.
⑧ When the ‘Select Diagnostics Kit’ window appears, select

‘HBV Quantitative PCR Kit’.
23
⑨ After the selection, ‘Check Prep Information’ window will

be appeared. Change the Initial Prep Vol. and Elution Vol.
according to the prep protocol. (e.g. ExiPrep™ Dx Viral DNA
Kit: Initial Prep Vol.-400㎕, Elution Vol.-80㎕) IU Conversion
Constant for HBV is ‘0.167. If all information is correct, click
the OK button.
⑩ After changing the prep information, find the sample layout

map saved in step ⑤ and click the ‘Open’ button.
⑪ When the ‘Sample Assign View’ window appears, verify that

the physical locations of the strips within the instrument
match the layout map. If the layout and locations match,
click the ‘OK’ button.
24
* If you need to make changes to the plate layout, click ‘Edit’

and make the necessary changes. When you are finished
making changes, click the ‘Update’ button. A pop-up will ask
you ‘Do you save your changes to Excel File?’ If you want
to permanently save the plate layout changes, select ‘Yes(Y)’
and proceed to select the original ‘*.xls’ plate layout file.

Click ‘Yes(Y)’ when the software asks you if you want to
replace the original file.
⑫ When the ‘Quick Start…’ window appears, verify sample

layout (located within the ‘Plate’ tab) and PCR protocol
information (located within the ‘Protocol’ tab). Click ‘OK’ if
everything is correct.
25
⑬ To perform the reaction, click the ‘▶‘button located within

the top toolbar.
26
9. DATA ANALYSIS
9.1. Process of Data Analysis
① Data can be analyzed by opening the data file through the

‘ExiDiagnosis Analysis’ program, which icon is located on
the computer Desktop screen. For details on how to analyze
data using the data analysis program, refer to the instrument
Instruction Manual.
② Select File>Open and select the data file (.ex3) to analyze.

The data files save in ‘C:\ExiDiagData\GUEST’ folder.
③ Several tabs are displayed as shown below. The HBV

Quantitative PCR Kit tab is configured to be initially
displayed.
27
④ The HBV Quantitative PCR Kit tab will display the calculated
Ct, ΔRn values and validity decisions in spreadsheet format.
⑤ The experiment fluorescence graph can be viewed in the Flu.

Graph tab.
28
⑥ The STD Curve tab displays the standard curve derived from
the run standards.
⑦ The Well vs Ct tab has the distribution of samples by Ct

value.
29
⑧ The Flu. Graph, STD Curve, Well vs Ct tab images can be

saved as *.jpg files. Use the save image icon ( ) within the
toolbar to save the image.
⑨ The HBV Quantitative PCR Kit tab contents can be exported

as a spreadsheet (*.xls). Select the save spreadsheet icon
( ) within the toolbar to save the spreadsheet.
30
9.2. Analysis Results
Fig. 4) Amplification curves of the standards and internal positive

control included within the kit for HBV diagnosis
Fig. 5) Standard curve of the five standards
31
Table 1. Cut-off table for data analysis

S/W version 1.27.4
Type FAM (HBV) TAMRA (IPC) Result
Ct: 20~35 Relative

Detection Limit 50~108 IU/㎖
ΔRn: >2000 Quantity
Ct: 20~35
NTC - Valid
ΔRn: >2000
Ct: 30.5~35.5
Standard 1
ΔRn: >2500
Ct: 27~31.5 Valid
Standard 2 (more than
ΔRn: >2500
Ct: 24.5~28.5 Ct: 20~35 3
Standard 3
ΔRn: >2500 ΔRn: >2000 Standards
Ct: 20.5~25 should be
Standard 4
ΔRn: >2500
valid)
Ct: 18~22
Standard 5
ΔRn: >2500
Relative
Sample 50~108 IU/㎖ +
quantity
Not
Sample - +
Detected
Sample 50~108 IU/㎖ - Repeat
Sample - - Repeat
32
10. TROUBLESHOOTING
Comments and suggestions

There is no FAM fluorescence signal in unknown samples and
Positive Control/ Standards.
• Thermal cycle program error
☞ Confirm that the correct thermal cycle was
programmed.
See 8.2.3. Programming the Exicycler™ 96 Real-Time
If the Quantitative Thermal Block.
TAMRA (IPC) • Omission of reaction component(s)

fluorescence ☞ Review your reaction preparation procedure.
signal was See 8.2.2. PCR Preparation.
detected • incorrect appointment of reporter and quencher dyes
☞ Check the appointments of the reporter and
quencher dyes.
See 8.2.3. Programming the Exicycler™ 96 Real-Time
Quantitative Thermal Block.
• A problem may have occurred during nucleic acid

extraction.
☞ Review your viral nucleic acid procedure. Check
If the samples for purity and quantity.
TAMRA (IPC)
• The kit may have spoiled, due to bad storage or the
fluorescence
kit may be past its expiration.
signal is
undetectable ☞ Assess your storage conditions and review the
expiration date.
Repeat the assay with new reagents in duplicate.
See 5. Storage.
33
A weak signal is present in the unknowns and Positive Control/

Standards.
• The reaction mixture has been incorrectly made.

☞ Review your reaction preparation procedure.
See 8.2.2. PCR Preparation.
• Inhibition of PCR
☞ Review your nucleic acid extraction procedure. Check purified
nucleic acids for quantity and purity.
• Reagent degradation from repeated freeze/thaw cycles.
☞ Assess your storage conditions and remaining kit components.
• The Kit has passed its expiration date.
☞ Review the expiration date printed on the Kit.
A signal is present in the Non Template Control (NTC).
• Contamination may have occurred

☞ Review your workspace and assess for contamination.
Signals are inconsistent between samples.
• There may have been a pipetting error

☞ Review the pipetting technique and calibration.
• Cross contamination may have occurred
• There may have been contamination on the optical adhesive cover
☞ Review your workspace and assess for contamination.
34
11. REFERENCES
Mackay IM. (2004) Real-time PCR in the microbiology laboratory.

Clin. Microbiol. Infect. 10:190-212
Servoss JC and Friedman LS (2004) Serologic and molecular

diagnosis of hepatitis B virus. Clin. Liver Dis. 8:267-281.
Schaefer S. (2005) Hepatitis B virus: significance of genotypes. J.

Viral Hepat. 12:111-124
35
12. SYMBOLS
Temperature
Catalog Number
Limitation
Contains
Manufacturer
Sufficient for test
Caution, Consult
accompanying Batch code
documents
Expiration Date Do not reuse
36
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AccuPower®

HBV Quantitative PCR Kit

Quantitative test kit

AccuPower® HBV Quantitative PCR Kit

Version No.: 2.2 (2012-12)

Safety Warnings and Precautions

Please inquire BIONEER’s Customer Service Center to obtain a copy of

Warranty and Liability

All BIONEER products are manufactured and tested under strict

AccuPower® is a registered trademark of BIONEER Corporation,

FAM and TAMRA are trademarks of Applera Corporation.

Excel™ is a trademark of Microsoft Corporation.

Copyright © 2012 by Bioneer Corporation. All Rights Reserved.

The AccuPower® HBV Quantitative PCR Kit is a ready-to-use,

This kit is optimized for use with BIONEER’s Exicycler™ 96

The use of kit is only for qualified users trained to correctly

Please read the instructions contained within this manual

Hepatitis B Virus (HBV) is reported to be closely related to

HBV infection can be determined by detecting HBsAg (HBV

AccuPower® HBV Quantitative PCR Kit is designed to quantify

3. FEATURES AND PRINCIPLE OF THE TEST

3.1. Kit Format

AccuPower® HBV Quantitative PCR Kit is a ready-to-use

3.2. Sensitivity and Dynamic Range

The sensitivity and the dynamic range of AccuPower® HBV

3.3 Internal Positive Control (IPC)

AccuPower® HBV Quantitative PCR Kit employs Internal

4. CONTENTS OF THE KIT

HBV Premix 8 well-strip 12 x 8-well strips

Optical sealing film 1 sheet

AccuPower® HBV Quantitative PCR Kit should

6. REQUIRED MATERIALS AND EQUIPMENT

- Disposable powder-free gloves

- Vortex and microcentrifuge capable of handling 8-well

- Store all kit components at -22 ~ -18℃.

8.1. Preparation: Specimen Collection, Storage, and

Caution: All samples should be treated as potential

8.1.1. Specimen Collection

AccuPower® HBV Quantitative PCR Kit is optimized for DNA

8.1.2. Sample Storage

8.1.3. Sample Transport

All samples should be transported in a shatterproof

8.1.4. Interfering Substances

Heparin (≥ 10 IU/㎖) is a known inhibitor of PCR. Samples

8.2. Work Flow Schema

8.2.1. Nucleic Acid Extraction

HBV DNA should be purified from raw clinical samples

* According to the extraction protocol or kit, real-time PCR

Products Cat. No. Platform

AccuPrep® Viral DNA Extraction Kit K-3511 Manual

ExiPrepTM Dx Viral DNA Kit K-4472 ExiPrepTM 16 Dx

8.2.2. PCR Preparation

① The appropriate number of premix tubes, controls and

② Briefly vortex and spin-down all reagents, controls and

③ Make a premix reconstitution mixture by combining 44 μl of

④ Add 45 μl of the premix reconstitution mixture to all premix

⑤ For the tubes that will be placed in A1 add 5 μl of DEPC DW

⑥ Move the tubes to a physically separated location before

⑦ Add 5 μl of nucleic acid extracts to the remaining tubes,

⑧ Cut the supplied Optical Film to size with scissors or a razor

spindown / 20 sec. hard vortex.

⑩ While the ExiSpin™ is operating, you can start the

⑪ After the ExiSpin™ cycling is complete, we recommend

8.2.3. Programming the Exicycler™ 96 Real-Time