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 TB-MAC Detection Kit hand Book


The TB-MAC detection Kit is a research reagent intended for detection of Mycobacterium

Tuberuculosis complex and M. avium complex (MAC) genome .

This product uses DNA chromatography strip : C-Pas ( GB6), which is produced by TBA , co., Ltd . Patent -Licences by NGK insulators, Ltd.

After PCR , we can  visually judge the existence of the target gene with DNA Chromatography method.

This kit is optimized for 10μl of PCR system .

2.Target Mycobacterium 

Target :                                                                                               Detection Line

Internarl control :                                                                                Tag 6

MAC                                                                                                        Tag 4

(M.avium, M.Intracellular, M.Chimera,M.Genavense.                Tag 4

M.Lentiiflavum, M.intermedium, M.arosiense,                             Tag 4

M.Colombiense,M.Florentinum, M.mantenii                                 Tag 4

M.Stomatepiae,M.Vulneris , M.youngonense                                 Tag 4

M.tuberculosis complex                                                                        Tag2

(M.tuberuculosis, M. Vulneris , M . Youngone nse)                        Tag2



Primer Soulution


Latex Solution

Dilution buffer

Low-Profile PCR tube

4.Other Necessary things

4-1 Reagents

  • Premix PCR Enzyme
  • Recommendation : SYBR Premix Ex Taq ) Tli RNaseH plus)(Takara RR20)
  • Quick TaQHS

4-2 Instruments

  • Gene amplification machine .
  • Recommendation:Quick Bath ) Thermo Gen QM-0104A)
  • Vortex Mixer
  • Micropipettes


TB-MAC detection Kit Hand Book 


5-1.Preparation of DNA solution

Please extract DNA from Samples .

5-2. Gene amplification by PCR

1)Prepare reaction mixture.

Add the following premix PCR solution to PCR tubes.

Premix PCR solution   5.0 μl

Primer Solution           2.5  μl

Total Volume                 7.5 μl

2)Add 205 μl of sample DNA solution ( Total 10 μl).

3)Amplify nucleic acids by PCR.

Be sure to close the PCR caps to avoid liquid leakage and evaporation during PCR. Then, Perform PCR under the following condition using Quick Bath.

PCR condition ( prese program No. SS65)

97°C 2 Min

97°C 5 sec

65 °C  10 sec

5-4.Notices in use of DNA chromatography strips : C-PAS(GB6)

Be sure to hold a part of absorbing pad of C-PAS) GB6) with dried hands in use .

Touching the parts other than absorbing pad and holding with me hands may result in and insufficient reaction.



TB-MAC detection Kit Hand Book



  • Absorbing Pad

Be sure to hold a part of absorbing pad of C-PAS(GB6) in use .

  • Position Marker

The position of emerging lines is discriminated by these lines.Position makers are always appeared with out without reaction.

  • Detection Line

Blue positive lines that exist before reaction temporary disappear when the solution passes through.

After that , reaction line appears again depending on the reaction .

Blue lines before use are gradually faded , but these changes don affect the reaction .

5-4.Reaction Using DNA chromatography Method

1)Take out Latex solution and Dilution buffer from refrigerator and keep at room temperature.

2)Prepare the Latex-Working solution by dilution the Latex Solution 1:10 in dilution Buffer.

Mix the solution uniformly with vortex mixer  by.

3)Open the caps of PCR tubes after PCR and add 11 μl  of the latex working solution . Mix the solution uniformly by pipetting.

4)Insert C-PAS (GB6)in tube of step “3)”.Top opposite of absorbing pad must be inserted tubes.

5)Leave them at room temperature for 10 minutes for developing . Perform the reaction at room temperature (20 ~30°C).

A low temperature causes flash – positive detentions.

5-5.Judgement of the results 

Negative : When no target gene exists in sample , the blue line at Tag 6(IC) line appears.

Positive :When target gene exists in sample , the blue line at detection line appears.
DNA=Strip -new-innovation-Japtec

IC line Often becomes faint when target gene is amplified.



Cocltail No.70B:TB-MAC Detection Kit

Species range covered with TB-MAC Detection Kit



Two Groups

M.Tuberculosis Group

M.avium Complex (MAC Complex)

Special Range




M.intermedium, M.genavens,






*Data Confirmed with 157 type strains of the genus Mycobacterium and 135 clinical mycobacterial strains including 53M.tuberculosis strains.

PCR amplicons are visualized with Blue Latex and DNA-Strip Chromatography.
All.M.Tuberculosis group strains make a single band on No2. position of DNA - Strip,
All M.avium complex strains make a single line on No.4 position of DNA strip .

Qick Mobile/ Quick Bath for Rapid Amplification of TB-MAC

  • Quick Mobile /Quick Bath( QM/QB) are designed to amplify targets within 25 minutes.
  • Quick Operation with per-installed Program Card. Just push appropriate program No.SS65 to Start.
  • QM/QB are applicable for outdoor use with 3 power supply systems of your choice .


Quick Bath (QB)

Low profile Tube:

16Well model (4.8kg)


Program Compatible>>>

Quick Mobile (QM)

Low profile Tube:

4 Well model (1.7 kg)




DNA-Strip Chromatography to identify TB-MAC Amplicon

  • After 25 Min Amplification, Add 10Ul of Blue Latex and plug DNA strip for Chromatography.
  • After 5-10 Minutes , Amplicon captured to DNA-Strip is Visualized as a solid blue line on a DNA-Strip .

DNA-Strip Identify

DNA-Chromato Technology is more Sensitive than Electrophoresis

M: Size Maker

  1. 1ng DNA
  2. 100pg DNA
  3. 10pg DNA
  4. 1pg DNA

N:Negative Control

  1. 5:100fg DNA
  2. 6:10fg DNA
  3. 7:1fg DNA

DNA-Chromatography have a potential to differentiate 5 different amplicons

DNA-Chromatography have potential -Japtec

Tow Different PCR tube Preparation 

A Liquid type( Low Profile PCR tube):48 tests

Two Type ( low Pfofile PCR Tube)


Example.Specimen Preparation

  1. Treat Sputum with standard NALC-Alkaline method.
  2. Centrifuge at 12,000gr for 5 minutes to precipitate cells.
  3. Discard the supernatant and add 200 Ul of distilled water.
  4. Vortex the tube and transfer the content to an MORA bead tube ( AMR products)
  5. Shake the tube whit Disrupted Jeanie for 2 min .
  6. Centrifuge the tube so spin down the solution at 2,500 rpm for a few second.
  7. Transfer 2.5 μl ( liquid primer) or 5.0  μl (prime dried tuble) into a PCR tuble for cocktail PCR.

kalin-Specimen Preoaration


Protocol B. Simple Boiling Method

  1. Collect sputum and mix with 2 fold dilution buffer .
  2. Transfer 200 Ul of the mixture into a MORE bead tube (AMR Product) And boil the Mixture at 100 c for 10 minutes.
  3. Vortex the tube and briefly centrifuge the tube.
  4. Transfer 2.5μl( liquid primer) or 0.5 μl ( primer dried tube) into a PCR tube for Cocktail PCR .


PCR and Chromatography Kit

Product features :

Simple genetic test kit

Easy · PCR chromatographic series product is a quick and simple genetic test reagent kit which is a combination of judgment for DNA chromatography with the tube to which the primer is dry-filled.

 Cocktail primer to match the inspection purpose

Pluralities of target genes will be amplified in a single reaction tube with a cocktail primer were mixed from 1 to select the six types of specific primers in a unique approach to each inspection purposes.

 Ready to use

Primer is dry-filled in the 8-connected PCR tube (Low profile).
Just add the enzyme and the template; it can be set directly to PCR equipment.
(. Note; limited to the Low profile tube Compatible models recommended equipment:

Saving the enzyme cost in 10μL reaction

This series product, since all have been optimized in 10μL reaction system, it can reduce the cost of the PCR enzyme.

Easy decision in DNA chromatography

After completion of the PCR reaction, you can visually determine the result only by inserting the DNA chromatographic added to the developing solution.
Also expensive real-time PCR instrument does not need cumbersome electrophoresis.

If you use the recommended PCR equipment (Bath), you can check in just 40 minutes until the determination from the PCR.

Building a wide variety of test kits series

A combination of specific primer of our ownership, can respond to a variety of inspection test kit series has built a.

Work flow of easy · PCR chromatographic

1. Preparation of the reaction tube 2. Preparation of the reaction solution 3.PCR reaction 4. The addition of a developing solution
Cut the required number with a pair of scissors or the like from the 8-connected PCR tubes that are dry-filled primer. In addition to a nucleic acid 5μL and XPCR enzyme 5μL to prepare a reaction solution. Set in a PCR instrument of 10μL reaction correspondence, performing PCR reaction. Add latex developing solution to the tube after the PCR reaction.
5. chromatography and reaction 6. result judgment Reaction example
Insert the chromatographic strip into a tube, and reacted for 5 to 10 minutes. It determines the presence or absence of each target gene by visual inspection. DNA chromatographic model Judgment example
Position3 positive

Product Specifications

Constitution Drying primer containing 8 consolidated PCR tube
DNA chromatographic strip
latex deployment solution
Reaction amount 48 reaction minute
STORAGE The cold storage (8 ℃ or less from the 2 ℃)