Data sheet NA-480 SALIVA DNA

Bio-Medical Department

GENE PREP STAR / SALIVA DNA Protocol

Automated extraction of gDNA fromOragene® saliva samples
GENE PREP STAR NA-480 / NA-480 PLUS (KURABO) is an automated DNA extraction systemthat can purify up to thirty samples. The purpose of this study is to determine the DNA
purification performance of the GENE PREP STAR NA-480 / NA-480 PLUS with saliva samples
collected with the Oragene® kit.

Saliva sample withOragene®

Sample tube
(NT-2100S)

NA-480 / NA-480 PLUS
Automated extraction system

DNA Collection tube
(NT-2100S)

TransferDNA Storage tube

Transfer

Automated
NA-480

ProcessNA-480 PLUS (ID tracking / LIMS mode)

Materials and methods
sample / volumeCollection

Saliva collected with Oragene® / 2 ml
Incubate saliva sample / Oragene® tube as below conditions.
50oC Water bath: >1 hour*
50oC Air incubator: >2 hours**Overnight incubation is acceptable.

Extraction system
ProtocolReagent kit
Consumables
Purification method

Automated DNA isolation system GENE PREP STAR NA-480 / NA-480 PLUS
SALIVA DNA (NA-480) / SALIVA+ 2ml (NA-480 PLUS)
NR-130, PR-4050, PR-5050
Sample tube : NT-2100S, 1.2mL Tip :T-1100,
Protein denaturation with non-phenol-based reagents
DNA precipitation by salting out

Process

Step1.

Protein denaturation & removal

Step2.

Precipitate DNA

Step3.

Wash DNA

Step4.

Eluate DNA (Standard elution volume: 500 μl)

URL; http://www.kurabo.co.jp/bio/Data sheet NA-480 SALIVA DNA

Bio-Medical Department

WorkflowSample tube

Add P Buffer
Saliva/ Oragene®
tubes

Transfer

Mix

centriferuge

Step1. Protein denaturation & removal

DNA tube

Add PR-4050

Add E Buffer

Add PR-5050

Waste

Waste

Mix

乾燥Mix

centricentri

Step2. Precipitate DNA

乾燥centrifuge

Step3. Wash DNA

Processing time

Heating & Mix

Step4. Elute DNA

10 sample: approx. 1.5 hours
20 sample: approx. 1.7 hours
30 sample: approx. 1.9 hours

AnalysisYield and Purity

Using the spectrophotometer NanoDrop (Thermo Scientific), measure the- Spectrophotometer

absorbance spectrum of 230 nm (A230), 260 nm (A260), 280 nm (A280), and320 nm (A320) of each isolates. Yield and purity are calculated as below.

PCR amplification
efficiency

Real-time PCR analysis was performed under the following conditions toconfirm the amplification efficiency of the product.
System : Quick Real-time PCR System Model GF-Q150 (KURABO)
Target gene : Human GAPDH (Amplicon size: 452 bp)
PCR Conditions：
Step

Temp&time

CyclesPre-denaturation

95 oC, 30s

1

denaturation

95 oC,

5s

annealing

60 oC,

10s

extension

72 oC,

10s

melt analysis

95 oC,

5s

40

1

URL; http://www.kurabo.co.jp/bio/Data sheet NA-480 SALIVA DNA

Bio-Medical Department

Results
Yield and Purity
Donor

PCR amplification
efficiency

Purity (A260/280)

Yield (μg)

A

1.824

73.6

B1.84335.0

C

1.717

7.2

D

1.764

173.3

E

1.831

19.6

F

1.734

91.6

G

1.812

37.7

H

1.739

145.2

I

1.850

35.7

J

1.778

37.0

K

1.837

21.1

L

1.873

13.7

M

1.785

106.5

N

1.830

71.6

O

1.804

77.2

Confirmed the amplification of the target gene Human GAPDH
Ct value：18-19*template : 10 ng

Note
Please note that in case of purifying DNA from human saliva, the results of purification may differ
significantly depending on the storage condition of the sample and individual differences.

Each product names written in this data sheet areregistered trademarks of each company

KURABO INDUSTRIES LTD.
14-30, Shimokida-cho, Neyagawa, Osaka 572-0823, Japan
KURABO Advanced Technology Center 2F
TEL: +81-72-820-3079
FAX: +81-72-820-3095

URL; http://www.kurabo.co.jp/bio/