F A S T Y P E

HLA -DNA SSP

TYPING SYSTEM
User’s Guide
For Class I HLA-ABC & Class II DR, DQ
Low, Intermediate and High Resolution

BIO•SYNTHESIS
INCORPORATED Copyright © 2000-2001 Bio-Synthesis, Inc., All Rights Reserved
 FASTYPETM HLA-DNA SSP TYPING SYSTEM PROTOCOL

1. PRIMARY FEATURES OF THE FASTYPETM HLA-DNA SSP TYPING SYSTEM:

1.1. SIMPLE HLA DNA TYPING

All primers, dNTPs, and buffers are pre-aliquot and lyophilized in high-quality PCR thermal cycler plates.

1.2. FAST PCR REACTION SETUP

FastypeTM HLA-DNA SSP Typing System Kits reduces the preparation time to less then 10 minutes.

1.3. ENHANCED STABILITY

Lyophilized primers, dNTPs and PCR buffer are more stable than in liquid form under ambient condition.

1.4. EASY CONTAMINATION DETECTION

Built-in contamination control primer pair(s) are designed to detect contaminations of genomic DNA and to detect carry-over PCR
end products.

1.5. UNIVERSAL PCR PROTOCOL

The PCR Protocol for all FastypeTM HLA-DNA SSP Typing System is kept the same regardless of the kit type.

1.6 UNION INTERNAL CONTROL BAND SIZE

The internal control band is kept the same at 410bp for all FastypeTM HLA-DNA SSP Typing Kits to reduce the confusion of positive
band versus internal control band.

2. PACKAGE CONTENTS, SHIPPING AND STORAGE CONDITIONS

SHIPPING CONDITION:
Ambient conditions/ room temperature.
PACKAGE CONTENTS STORAGE SHELF LIFE

• FASTYPETM KIT -20OC 1 year or as specified on the

package label
• REAGENT E -20OC 1 year

• 10X DYE -4OC 1 year

• 8-WELL STRIP CAPS AMBIENT CONDITIONS

3. TYPING PROTOCOL

3.1 COMPONENTS OF FASTYPETM HLA-DNA SSP TYPING SYSTEM KIT:

a. Allele-specific or group-specific primer pair(s)
b. Internal control primer pair specific for human G3PDH gene
c. dNTPs
d. PCR buffer
3.2 MATERIALS
(See Appendix B for required material and instrument)
3.3 SETTING UP PCR
(See Figure 3.1 for PCR parameters)

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FOR RESEARCH USE ONLY
 FASTYPETM HLA-DNA SSP TYPING SYSTEM PROTOCOL

(BELOW STEPS ARE TO BE PERFORMED IN PRE-PCR AMPLIFICATION ROOM )

3.4. PREPARING REAGENT
3.4.1. Obtain a 1.5 ml- microcentrifuge tube.
3.4.2. Combine equivalent volumes of Reagent E and Taq Polymerase (5u/ul) into an 1:1 (v/v) mixture.
3.4.3. Incorporate the mixture in room temperature for 5 minutes. Store the remaining at -20oC.
3.5. PREPARING DNA MASTER MIX
3.5.1. Remove FastypeTM SSP kits from -20oC. Bring to room temperature. Secure on a 96-well microtiter plate holder.
Note: You can mix and match different FastypeTM Typing Kits to maximize your typing needs.
3.5.2. Bring the Cresol red Dye (10X) to room temperature. Vortex thoroughly before using.
Note: For short term storage, please kept at 4oC.
3.5.3. Obtain a 1.5ml microcentrifuge tube. Prepare Master Mix (See Appendix A for Master Mix formulation)
3.5.5. Vortex the master mix thoroughly, spin down if necessary. Pipette 10ul into each well (or tubes).
Important: Upon adding the master mix, the contents inside should turn into a bright pink color solution. If the solution
turns yellow, your sample may be too acidic and may effect PCR amplification.

3.6. PREPARING FOR CONTAMINATION CONTROL DETECTORS

3.6.1. Prepare the following contents in built-in contamination control well (or tubes):
Sterile ddi H2O 9 ∝l
10X dye solution 1 ∝l
Taq/Reagent E 0.1 ∝l
The final volume should add up to 10∝l.
3.8. SEALING
3.8.1. Gently tap the wall of the PCR tube to settle master mix to the bottom of the wells (or tubes).
3.8.2. Add one drop (~15∝l) of mineral oil to each tube (optional).
3.8.3. Seal the reaction tubes with the caps provided or with a PCR mat.
3.9. SECURE ON A THERMAL CYCLER AND BEGIN PCR CYCLE.

FIGURE 3.1 PCR PARAMETER

Thermal Cycler Protocol

Program the thermal cycler with the following parameters:

Tm Time Cycle No. Linked to

Program 1 96oC 1 min 1 Program 2

Program 2 Segment 1 94oC 20 sec Program 3

Segment 2 65oC 60 sec 10

Program 3 Segment 1 94oC 20 sec Program 4

Segment 2 61oC 50 sec 20

Segment 3 72oC 30 sec

Program 4 72oC 5 min 1

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FOR RESEARCH USE ONLY
 FASTYPETM HLA-DNA SSP TYPING SYSTEM PROTOCOL

4. AGAROSE GEL ELECTROPHORESIS

THE AMPLIFIED BANDS CAN BE VISUALIZED BY SUBMARINE AGAROSE GEL ELECTROPHORESIS.

4.1. PREPARING 2% (W/V) AGAROSE GEL

4.1.1. Dissolve 2.0 grams of electrophoresis grade agarose powder in 100 ml of 0.5X TBE buffer. (See Appendix B. for buffer
preparation.)
4.1.2. Melt the agarose powder completely by boiling in a microwave oven or on a heating apparatus. Stir constantly. If
evaporation occurs, replenish with ddi H2O.

4.2 CASTING GEL

4.2.1. Cool the heated agarose gel to ~60oC
4.2.2. Add at least 2 ul of ethidium bromide (10 mg/ml ) to the heated agarose. Stir until the it is thoroughly incorporated.
4.2.3. On a balanced surface, set up a gel plate with combs containing a minimum of 9 wells per row.
4.2.4. Cast a 5mm thick gel on the plate.
4.2.5. Allow the gel to settle.

(BELOW STEPS ARE TO BE PERFORMED IN POST-PCR AMPLIFICATION ROOM)

4.3 GEL ELECTROPHORESIS
4.3.1. Submerge the gel in 0.5X TBE buffer in a gel box.
4.3.2. Gently remove the caps to avoid splashing of PCR products.
4.3.3. Start pipette from the bottom-left of the kit and proceed to the right and upward. (Refer to “Fastype Kit Configuration”
for details)
4.3.4. Load a minimum of 8 ∝l into each well on the gel.
Note: The PCR products are stained with cresol red. There is no need to load additional “loading buffer” or dye.
4.3.5. (Optional) Reserve the first well on each row for molecular weight standards, in increments of 100bp from 50 to 1000bp.
4.3.6. Connect the electric leads and turn on the power supply (115V AC ). Electrophoresis for ~ twenty (20) minutes, or until
two thirds (2/3) of the lane.
4.3.8. Turn off the power supply, and remove the gel from the gel box.

4.4 VISUALIZING RESULTS

4.4.1. Transfer the gel onto a UV transilluminator.
4.4.2. Document the result by photography.
4.4.3. Use the corresponding typing sheet and table to interpret results.

5. INTERPRETATION OF THE RESULT

Attention: Due to continuous updating, the typing sheet and specificity table may vary from lot to lot. Upon receiving, please
confirm the lot number on the package label matches the typing sheet and specificity table provided. Should you have any
questions, please contact our Technical Support at 1-800-227-0627.

5.1. Refer to Typing Sheet and Specificity Table for product sizes and primer information.
5.2. All Internal control band has a size of 410 bp. The intensity of the internal control PCR product may vary due to the
presence of positive amplifications.
5.3. If the PCR products have bands other than the predicted size, disregard the band for it may be none specific or primer-
dimer artifacts.
5.4. The presence of a 95bp band in contamination control lane implies a contamination of Post-PCR Product. The presence
of a 410bp band alone or accompanied by a 95bp band implies a contamination of genomic DNA.

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FOR RESEARCH USE ONLY
 TROUBLE SHOOTING AND TECHNICAL GUIDE

6. TROUBLE SHOOTING GUIDE

PROBLEMS POSSIBLE CAUSE SUGGESTIONS
1. The control and specific bands are a. Concentration of DNA sample is too a. Check DNA quality and concentration
weak. low. (see Technical Guide).
b. Degradation of sample DNA. b. Re-purify the sample DNA and dissolve
c. Inhibitors for Taq polymerase present in it in ddiH2O.
the DNA sample.

2. Heavy smear of bands or dim specific Template DNA is in excess. Check DNA concentration and adjust
bands accordingly.

3. Missing internal control bands in Repeat typing with another good quality
one or several lanes DNA sample.

4. False negative of a specific band
while the internal control appears
normal.
Contamination with previously amplified
5. More than two specific alleles are PCR products or with other DNA samples
detected during the DNA extraction or PCR prepara-
tion.
6. Ambiguous results
a. The electrophoresis buffer might
7. Blurred bands become too hot due to high voltage.
b. Wrong buffer composition.
c. Old buffer
Repeat typing with a control DNA sample
which has the same specific allele.
a. Separate the pre-PCR room from the
post-PCR room.
b. Kept a different lab coat between rooms.
c. Clean the working area with 10% bleach.
d. Change protective gloves frequently,
especially transfer from post- PCR room to
pre-PCR room.
Report to Technical Support
a. Lower the voltage during gel
electrophoresis.
b. Use the recommended 0.5 x TBE buffer.

7. TECHNICAL GUIDE

7-1. DNA Quality

For optimal results with the FastypeTM DNA typing system, the quality of sample DNA is critical. Good quality DNA means the 260/280
ratio of OD is higher than 1.6 and the major portion of DNA should run higher than 9.4 kb on an agarose gel. Heavy degradation of
genomic DNA or a 260/280 ratio lower than 1.5 requires DNA re-extraction. For best result, dilute sample DNA in ddiH2O.

7-2. DNA Quantity

The quantity of sample DNA should be 25- 50 ng per reaction. Excess of sample DNA (100 ng or more) can cause decline of specific
PCR products. An intense smear of high molecular DNA is an indication of excess of sample DNA. RNA contamination can cause
higher readings of DNA concentration with a spectrophotometer. If PCR reactions fail, it is always recommended to check the quantity
and quality of sample DNA by running sample DNA in 1% agarose and compare it with a series of standard Lambda DNA marker.

7-3. Taq polymerase

FastypeTM kits have been intensively tested with the AmpliTaq DNA polymerase from Perkin-Elmer. We recommend that users use
AmpliTaq DNA polymerase as the first choice. Do not use Tth DNA polymerase.

7-4. Amplification Procedure

At the end of PCR, examine the degree of evaporation and condensation of PCR reaction mixture. If there is more than 20% volume
loss, you should overlay the PCR reaction mixture with mineral oil. It is also a good practice to maintain QC records on the heating lid.
If the temperature of the heating lid is not high enough, it will cause condensation problems on the lid.

7-5. Thermal cycler

Perkin-Elmer 9600 and 9700 thermal cyclers are recommended for Fastype HLA Class I and Class II typings.

IF PROBLEMS STILL PERSIST, CALL THE TECHNICAL SUPPORT AT 1-800-227-0627

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FOR RESEARCH USE ONLY
 FASTYPETM KIT CONFIGURATION
FASTYPE KIT CONFIGURATION
Kit Type Kit Type

HLA-A (cat. 20-0290-FA) DRB1*01 (cat. 20-1201-F)

17 18 19 20 21 22 23 CC Color Code: Purple 1 2 3 4 5 6 7
Color Code: Blue CC
No. of wells: 8
No. of wells: 24 9 10 11 12 13 14 15 16
1 2 3 4 5 6 7 8 DRB1* 03 (cat. 20-1203-F)
9 10 11 12 13 cc
Color Code: White
HLA-B (cat. 20-0290-FB) No. of wells: 14 1 2 3 4 5 6 7 8
Color Code: White 41 42 43 44 45 46 47
No. of wells: 47 33 34 35 36 37 38 39 40 DRB1*04 (cat. 20-1204-F)
9 10 11 12 13 14 15 16
Color Code: Blue
25 26 27 28 29 30 31 32 No. of wells: 16 1 2 3 4 5 6 7 8
17 18 19 20 21 22 23 24 DRB1* 07 (2 tests per strip)
9 10 11 12 13 14 15 16 (cat. 20-1207-F) 1 2 3 CC 1 2 3 CC
Color Code: Red
1 2 3 4 5 6 7 8
DRB1*08 (cat. 20-1208-F)
Color Code: Yellow 9 10 11 12 13 14 15 16
HLA-C (cat. 20-0290-FC) 17 18 19 20 21 22 23 No. of wells:16
CC 1 2 3 4 5 6 7 8
Color Code: Yellow
No. of wells: 24 9 10 11 12 13 14 15 16 DRB1*11 (cat. 20-1211-F)
Color Code: Red 9 10 11 12 13 14 15 16
1 2 3 4 5 6 7 8 No. of wells: 16
1 2 3 4 5 6 7 8
DR Low (cat. 20-0290-FR) DRB1*12 (cat. 20-1212-F)
17 18 19 20 21 22 23 CC Color Code: Red 1
Color Code: Red 2 3 4 5 6 7 8
No. of wells: 8
No. of wells: 24 9 10 11 12 13 14 15 16
1 2 3 4 5 6 7 8 DRB1*13 (cat. 20-1213-F) 9 10 11 12 13 14 15 16
Color Code: White
DQ Low ( cat. 20-0290-FQ) No. of wells: 16 1 2 3 4 5 6 7 8
Color Code: Green 1 2 3 4 5 6 7 8 DRB1*14 (cat. 20-1214-F)
No. of wells: 8 Color Code: Blue 9 10 11 12 13 14 15 16
No. of wells: 16
1 2 3 4 5 6 7 8
DQ High (2000) 25 26 27 28 29 30 31 32
DRB1*15 (cat. 20-1215-F)
(cat. 20-0290-FQH)
17 18 19 20 21 22 23 24 Color Code: Purple 1 2 3 4 5 6 7 8
Color Code: Green No. of wells: 8
No. of wells: 32 9 10 11 12 13 14 15 16
DRB1*16 (cat. 20-1216-F) 1 2 3 4 5 6 7 8
1 2 3 4 5 6 7 8 Color Code: Red
No. of wells: 8
DQ Intermediate
17 18 19 20 21 22 23 24 9 10 11 12 CC
(Previously DQ High) DRB3 (cat. 20-12B3-F)
(cat. 20-0290-FQI) 9 10 11 12 13 14 15 16 Color Code: Yellow 1 2 3 4 5 6 7 8
No. of wells: 13
Color Code: Green
1 2 3 4 5 6 7 8
No. of wells: 24
DRB4 (cat. 20-12B4-F) 1 2 3 4 5 6 7 8
Color Code: Green
9 10 11 12 13 14 15 16 No.of wells: 8
DQA (cat. 20-0290-FQA)
Color Code: Green 1 2 3 4 5 6 7 8 9 10 11 12 CC
DRB5 (cat. 20-12B5-F)
No. of wells: 24 Color Code: White
1 2 3 4 5 6 7 8
No.of wells: 13

NOTE:
1. “CC” refers to built-in contamination control.
2. The “COLOR CODE” refers to the plate color, the ring color if it is a
8-well strip, or unless specified otherwise.

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FOR RESEARCH USE ONLY
 MASTER MIX GUIDE Appendix A
T aq + Re ag e n t
S a m p le D N A C re s o l Re d
d d iH 2 O E m ix t u r e T o t a l V o lu m e
( ~ 5 0 n g /u l) n Dye (10X)
(5 u n its /u l)
(u l) (u l) (u l) (u l) (u l)
1 1 8 1 0 .1 1 0 .1
2 2 1 6 2 0 .2 2 0 .2
3 3 2 4 3 0 .3 3 0 .3
4 4 3 2 4 0 .4 4 0 .4
5 5 4 0 5 0 .5 5 0 .5
6 6 4 8 6 0 .6 6 0 .6
7 7 5 6 7 0 .7 7 0 .7
8 8 6 4 8 0 .8 8 0 .8

9 1 0 8 0 1 0 1 1 0 1
1 0 1 1 8 8 1 1 1 .1 1 1 1 .1
1 1 1 2 9 6 1 2 1 .2 1 2 1 .2
1 2 1 3 1 0 4 1 3 1 .3 1 3 1 .3
1 3 1 4 1 1 2 1 4 1 .4 1 4 1 .4
1 4 1 5 1 2 0 1 5 1 .5 1 5 1 .5
1 5 1 6 1 2 8 1 6 1 .6 1 6 1 .6
1 6 1 7 1 3 6 1 7 1 .7 1 7 1 .7

1 7 1 9 1 5 2 1 9 1 .9 1 9 1 .9
1 8 2 0 1 6 0 2 0 2 2 0 2
1 9 2 1 1 6 8 2 1 2 .1 2 1 2 .1
2 0 2 2 1 7 6 2 2 2 .2 2 2 2 .2
2 1 2 3 1 8 4 2 3 2 .3 2 3 2 .3
2 2 2 4 1 9 2 2 4 2 .4 2 4 2 .4
2 3 2 5 2 0 0 2 5 2 .5 2 5 2 .5
2 4 2 6 2 0 8 2 6 2 .6 2 6 2 .6

2 5 2 7 2 1 6 2 7 2 .7 2 7 2 .7
2 6 2 8 2 2 4 2 8 2 .8 2 8 2 .8
2 7 2 9 2 3 2 2 9 2 .9 2 9 2 .9
2 8 3 0 2 4 0 3 0 3 3 0 3
2 9 3 1 2 4 8 3 1 3 .1 3 1 3 .1
3 0 3 2 2 5 6 3 2 3 .2 3 2 3 .2
3 1 3 3 2 6 4 3 3 3 .3 3 3 3 .3
3 2 3 4 2 7 2 3 4 3 .4 3 4 3 .4

3 3 3 6 2 8 8 3 6 3 .6 3 6 3 .6
3 4 3 7 2 9 6 3 7 3 .7 3 7 3 .7
3 5 3 8 3 0 4 3 8 3 .8 3 8 3 .8
3 6 3 9 3 1 2 3 9 3 .9 3 9 3 .9
3 7 4 0 3 2 0 4 0 4 4 0 4
3 8 4 1 3 2 8 4 1 4 .1 4 1 4 .1
3 9 4 2 3 3 6 4 2 4 .2 4 2 4 .2
4 0 4 3 3 4 4 4 3 4 .3 4 3 4 .3

4 1 4 4 3 5 2 4 4 4 .4 4 4 4 .4
4 2 4 5 3 6 0 4 5 4 .5 4 5 4 .5
4 3 4 6 3 6 8 4 6 4 .6 4 6 4 .6
4 4 4 7 3 7 6 4 7 4 .7 4 7 4 .7
4 5 4 8 3 8 4 4 8 4 .8 4 8 4 .8
4 6 4 9 3 9 2 4 9 4 .9 4 9 4 .9
4 7 5 0 4 0 0 5 0 5 5 0 5
4 8 5 1 4 0 8 5 1 5 .1 5 1 5 .1

5 0 5 4 4 3 2 5 4 5 .4 5 4 5 .4
5 2 5 6 4 4 8 5 6 5 .6 5 6 5 .6
5 4 5 8 4 6 4 5 8 5 .8 5 8 5 .8
5 6 6 0 4 8 0 6 0 6 6 0 6
5 8 6 2 4 9 6 6 2 6 .2 6 2 6 .2
6 0 6 4 5 1 2 6 4 6 .4 6 4 6 .4
6 2 6 6 5 2 8 6 6 6 .6 6 6 6 .6
6 4 6 8 5 4 4 6 8 6 .8 6 8 6 .8

6 6 7 1 5 6 8 7 1 7 .1 7 1 7 .1
6 8 7 3 5 8 4 7 3 7 .3 7 3 7 .3
7 0 7 5 6 0 0 7 5 7 .5 7 5 7 .5
7 2 7 7 6 1 6 7 7 7 .7 7 7 7 .7
7 4 7 9 6 3 2 7 9 7 .9 7 9 7 .9
7 6 8 1 6 4 8 8 1 8 .1 8 1 8 .1
7 8 8 3 6 6 4 8 3 8 .3 8 3 8 .3
8 0 8 5 6 8 0 8 5 8 .5 8 5 8 .5

8 2 8 8 7 0 4 8 8 8 .8 8 8 8 .8
8 4 9 0 7 2 0 9 0 9 9 0 9
8 6 9 2 7 3 6 9 2 9 .2 9 2 9 .2
8 8 9 4 7 5 2 9 4 9 .4 9 4 9 .4
9 0 9 6 7 6 8 9 6 9 .6 9 6 9 .6

9 2 9 8 7 8 4 9 8 9 .8 9 8 9 .8

9 4 1 0 0 8 0 0 1 0 0 1 0 1 0 1 0

9 6 1 0 2 8 1 6 1 0 2 1 0 .2 1 0 3 0 .2
Note: “n” is the number of reactions. To compensate for pipette variations, each master mix above ( where n > 16) is adjusted with 2 extra reaction to (n+2).
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 Appendix B

TABLE 1. INSTRUMENTS AND MATERIALS TABLE 2. ELECTROPHORECTIC SOLUTIONS

PCR Recommended
Amplification Suppliers 10X TBE Buffer
1L pH 8.0
Thermal Cycler Perkin-Elmer
GeneAmp PC Tris-base 108g
system 9600 or Boric Acid 55g
EDTA 14.8 g
9700
(Sodium Salt)
Taq Polymerase Dissolve all ingredients in ddiH2O and adjust
AmpliTaq DNA
the volume to 1L.
Polymerase
Filter the solution to remove particles.
Perkin-Elmer
Store in bottles at room temperature (25oC).
N801-0060
Sterile Water
0.5X TBE Buffer
Gel Electrophoresis
Dilute 500ml 10X TBE buffer to 10 L ddiH2O
Gel box Bio-TyperTM Store in ambient condition.
Electrophoresis
System
20-0180-25 / Ethidium Bromide Solution
20-0180-09 100 ml

Agarose Life Technologies Ethidium Bromide 1g

(electrophoresis
grade) Dissolve EtBr in 100ml ddiH2O
Stir for several hours until the dye has
dissolved completely.
Ethidium Bromide Sigma E-1510 Store in an amber bottle or wrap with
0.5X TBE Buffer aluminum foil to avoid light.
Store in ambient condition, up to 12 months
Microwave oven /
Heating Equipment
Stir Plate
WARNING!
Power Supply Ethidium bromide is a powerful mutagen! Handle with
(115V AC 50-60Hz) care.
UV Trans-illuminator
Polaroid camera/film
or Video camera/printer

PCR LICENSE
The use of polymerase chain reaction (PCR) for in-vitro diagnostic procedure requires a license. The PCR is
covered by United States Patents 4,683,195 and 4,683,202 and corresponding patent rights in other territories,
owned by F. Hoffmann-La Roche Inc.
License can be obtained from:
Director of Licensing
Roche Molecylar Systems
1145 Atlantic Avenue
Alameda, CA 94501 USA

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 REFERENCES

10. REFERENCES

1. Bidwell, J.H. Application of the Polymerase Chain Reaction to HLA Class II Typing, Vox Sang 63: 81-8. (1992)

2. Bunce, M., et. al. Phototyping: comprehensive DNA typing for HLA-A, B,C DRB1, DRB3, DRB4, DRB5 & DQB1by
PCR with 144 primer mixes utilizing sequence-specific primers (PCR-SSP). Tissue Antigens 46: 355-367. (1995)

3. Chen, R., et.al. High Resolution Typing of HLA-DR by PCR amplification with sequence -specific primers (PCR-SSP).
Human Immunology 22nd Annual ASHI Metti8ng Abstracts 49 7.4 #182:133 (1996)

4. Innis, M.A., et. al. PCR Protocols: A guide to methods and applications. Academic Press, San Diego, CA. 1990.

5. Olerup, O., et. al. HLA-DR typing by PCR amplification with sequence-specific primers (PCR-SSP) in 2 hours: An
alternative to serological DR typing in clinical practice including donor-recipient matching in cadaveric transplantation.
Tissue Antigens 39: 225-35. (1992)

6. Olerup, O., et. al. HLA-DQB1 and DQA1 typing by PCR amplification with sequence-specific primers (PCR-SSP) in 2
hours. Tissue Antigens 41: 119-134. (1993)

7. Robinson J, Malik A, Parham P, Bodmer JG, Marsh SGE: “IMGT/HLA database - a sequence database for the human
major histocompatibility complex.” Tissue Antigens (2000) 55 280-287
http://www.anthonynolan.org.uk/HIG/data.html

8. Saiki, R.K., et. al. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase, Science 238:
487-9. (1988)

9. Sambrook, J. et. al. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor,
N.Y., 1989.

10. Zetterquist, H. et. al. Identification of the HLA DRB1*04, -DRB1*07 and -DRB1*09 alleles by PCR amplification with
sequence-specific primers (PCR-SSP) in 2 hours. Human Immunology 34: 64-74. (1992)

LIMITED WARRANTY &TERMS

FastypeTM system products have a one year manufacturer warranty. This warranty is valid only to the original purchaser for one year
from the date of purchase. At Bio-Synthesis’ discretion, Bio-Synthesis’ obligation to the original purchaser is limited either the
replacement of the product without extra charge, or a total reimbursement. The purchaser is responsible for the retuning of any
defective items to Bio-Synthesis Incorporated. All shipment must be such that no damage is incurred. Claims for shipping damage
must be submitted to the transit carrier.

This warranty becomes null and void when any product(s) have been modified, repackaged or resold without Bio-Synthesis’ consent,
or when any product(s) have been subjected to misuse or mishandling.

Bio-Synthesis Incorporated will replace or refund to the original purchaser if the purchaser is dissatisfied with the overall performance
of FastypeTM system within the first thirty (30) days of purchase. If the purchaser has any other concern, please contact our service
support at 1800-227-0627.

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FOR RESEARCH USE ONLY