(1) Detection of Helicobacter pylori through PCR

 

The culture method has been used conventionally to detect Helicobacter pylori from

gastric biopsy specimens. Now the PCR method has been gradually recognized as an

effective alternative since it enables sensitive detection and results can be obtained in

around seven hours, whereas the culture method takes at least a week.

The performance of TaKaRa Taq and TaKaRa Ex Taq was compared in detection by

directly applying gastric biopsy specimens to PCR. The result showed that TaKaRa Ex

Taq could detect Helicobacter pylori from the specimen while TaKaRa Taq could not.

This indicates that TaKaRa Ex Taq minimizes the risk of false-negative results which can

occur using conventional Taq DNA polymerases, and it achieves rapid detection with

the same level of accuracy as the culture method.

 

 

Template:
DNA extracted from gastric biopsy

specimens collected

from the patients

with gastritis ulcer

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 (2) Amplification of human genomic DNA directly from human blood 

 

 Performance was compared between TaKaRa Ex Taq and TaKaRa Taq when human

blood samples which were treated with anticoagulants like sodium citrate and EDTA

were directly applied to PCR reaction as template. The target was the cytokeratin 19

region of human genomic DNA. In both cases using TaKaRa Ex Taq and TaKaRa

Taq, larger sample volume gave greater amplification inhibition by anticoagulants.

However, it was confirmed that TaKaRa Ex Taq is less affected by anticoagulants than

TaKaRa Taq.

Heparin is known to inhibit enzyme reactions relating to nucleic acids. However, the

figure confirms that the addition of heparinase (around 0.5 unit) into the reaction mixture

before PCR decreases the inhibition by heparin.

 

 

 

 

 

When blood samples treated with sodium citrate or EDTA were used as the template

 

 3% NuSieve 3:1 Agarose gel (BMA) Applied volume:
8

 

 

 

When blood sample treated with heparin was

applied as the template (with/without addition of heparinase)

 

 

 

 

 

(3) Amplification of extracted DNA using TaKaRa DEXPAT 

 

 Extracted DNA from 6 paraffin sections (ea. 2 x 10 ) using TaKaRa DEXPAT were used as the templates for PCR. In conjunction with TaKaRa Ex Taq accurate and highly sensitive PCR can be achieved. Extracted DNAs were stable at 4°C for at least one month.

Lane M:

pHY Marker (100 ng)

Target region:
-globin gene

Template:

Extracted DNA using TaKaRa DEXPAT

PCR:
50