Technology

Why is UV-light so dangerous?

IntroductionDetection of nucleic acid is still performed using light in the UV-range and ethidium bromide as nucleic acid stains. However, the degradation of nucleic acids by UV-light is well documented and can therefore be considered a nucleic acid destroying agent. Additionally, the UV-light is considered dangerous for the user. Overall, the effi ciency of downstream applications, such as sequencing, cloning, etc. can be signifi cantly reduced due to damaged DNA molecules. Harmless visible light has already been used to detect nucleic acids with its major limitation of only exciting green dyes. NIPPON Genetics EUROPE, known for its safe nucleic acid dyes MIDORIGreen, has developed a complete new approach for the detection of nucleic acid dyes able to excite all commercially available nucleic acid dyes.

DNA is able to absorb light in the UV-spectrum. Consequences are the formation of pyrimidine dimers; ssDNA and dsDNA breaks, DNA-DNA interstrand crosslinks and DNA-to-protein crosslinks.

Organisms are able to repair these DNA damages or suffer the consequences discussed above. DNA in agarose gels lack these repair mechanisms.

Additionally, the amount of DNA in a gel is much less when compared to a whole organism. Therefore DNA degradation caused by short-term UV light exposure was investigated:

After an electrophoretic run, the gel was exposed to UV-light for up to 60 seconds. The PCR product was subsequently isolated and cloned into a control vector, previously double digested with NdeI and HindIII, before being cloned and transformed into Escherichia coli DH5α (BL21) cells. The results are shown below:

The DNA exposed to UV-light was highly damaged, signifi cantly reducing the cloning effi ciency. After 60 seconds exposure, no colony was detected, indicating complete damage of DNA.

In 2009, blue light emitting diodes (LED) transilluminators were introduced. LED are semi-conductors able to transform electricity into light. The advantages are defi ned wavelengths, superb effi ciency and a huge average life expectancy of 50,000 hours. They emit light with a wavelength of 470 nm, which is after the UV-wavelength range.

The experiment described previously was repeated and the DNA was exposed to the blue LED light for the same amount of time:

As expected from the physical theory, the amount of CFU per plate did not have any signifi cant change after the exposure. The prevention of the DNA damage therefore vastly increased the cloning effi ciency.

Blue LED - First Solution

Problems with Blue LED

Although having the assets of safe light source, using blue LEDs also had a central drawback. Ethidium bromide, the most common DNA stain did only deliver poor results. The

excitation peak of ethidium bromide bound to DNA is between 480 and 525 nm.

As can be seen, the signal of ethidium bromide bound to DNA is very week and cannot be used. Green DNA dyes, such as MIDORIGreen dyes, deliver a much better signal with blue LEDs.

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NGFG09 NGFG12

FAS-V

Key Features• Very sensitive 2MPixel CCD-Camera (0.13 Lux)

• 10.4” Touchscreen display

• 26 cm x 21 cm Blue/Green LED Transilluminator

• 26 cm x 21 cm White LED Transilluminator

The FastGene® FAS-V is our most advanced imaging system. It is a standalone imaging system with an internal computer and a 10.4“ touchscreen. It contains our largest imaging area yet, for DNA as well as for protein imaging. The super sensitive CCD-Sensor is combined with a high-end parfocal lens, normally only found in microscope. The combination of our unique excitation technology with the high-end components deliver the best results with an easy to use system.

• Detection and documentation of DNA/RNA dyes

• Detection of GFP, eGFP, YFP, etc.

• Documentation of protein gels

• Documentation of Petri dishes & membranes

• On-side image editing

Applications

Image formats• Jpeg

• TIFF

• BMP

• PNG

Blue/Green LED TransilluminatorUnique Excitation Technology

• Enables the detection of red and green DNA dyes• Huge imaging area is 26 cm x 21 cm big • Superb uniform transillumination• 12 large LED arrays at each side

White LED Transilluminator• Detection of protein • Very large white LED table (26 cm x 21 cm)• Directly controlled by the touchscreen

White LED Room light• Documentation of membranes and Petri dishes• Two white LED arrays illuminate the imaging box

CCD-Sensor• CCD-Sensor size of 1/1.8“• Pixel size of 4.4 µm • Detection of very low signals of 0.13 Lux*• Images can be recorded in TIFF, JPEG, BMP and PNG format• 16 GB internal SSD storage or external USB-stick• Exposure time from 0.001 sec up to 30 sec• Manufactured in Japan

*Clarity of a normal day is between 20 000 and 100 000 Lux

Very bright parfocal lens - no more focusingThe lens of the FastGene® FAS-V is parfocal (preinstalled focus does not have to be readjusted when zooming in or out)• Huge aperture* of f/1.2• Aperture is regulated steplessly, • 6 x Zoom • Focal length of 12.5 mm to 75 mm

*opening of the lens iris

Optical Unit

Illumination

Comparison

The FAS V Imaging System has a highend Japanese CCD-sensor able to detect 0.13 Lux

2 MPixel UXGA CCD Sensor

10.4” Touchscreen Display

Intuitive Software

White LED room light

Blue/Green LED Transilluminator

White LED Transilluminator

The Blue/Green LED technology is able to excite red and green DNA dyes, e.g. MIDORIGreen, SYBR™ & EtBr

FAS-V

The imaging software takes 6 shots simultaneously, with different exposure times and includes an image editor

Ethidium Bromide MIDORIGreen Direct MIDORIGreen Advance

TouchscreenThe FastGene® FAS-V is controlled by a 10.4“ touchscreen

• All three light sources can be activated or deactivated by the touchscreen.

• Exposure time and gain adjustment can be adjusted using the slide function or by directly adding the desired value.

• Freezing the image

• Capturing the image

• Inverting live view or loaded image

Image EditingRecorded images can be directly edited

• Adjusting contrast

• Cropping the image

• Rotation of the image anti- and clockwise

• Mirroring vertically and horizontally

• Inverting black and white

ConnectionsStorage

The internal SSD-storage of the FastGene® FAS-V has a size of 16 GB. This can be expanded using an external USB-Stick. There are 3 USB ports to connect USB-sticks, mouse or a keyboard

NetworkThe FastGene® FAS-V can be directly connected to a network using the 100 MBits ethernet port

PrintingThe FastGene® FAS-V can be used with the thermal printer Mitsubishi P95D

FAS-V

Camera sensor type 2MPixel CCD - charge-coupled deviceSensor and image size 1/1.8” Sensor with 1600 x 1200 (UXGA)Mininmal illumination 0.13 LuxImage format TIFF, JPEG, BMP, PNGExposure time 0.001 to 30 secPixel size 4.4 x 4.4 µmAperture f / 1.2Focal distance 12.5 - 75 mm Zoom range 6 x zoom Filter Removable amber fi lter

Display 10.4” Color touchscreen Display resolution 800 x 600

Internal storage 16 GB SSDExternal storage USB 2.0Connections Ethernet (100Mb), USB 2.0, Thermoprinter Mitsubishi P95D supportedSoftware Touchscreen with image editor software

Light sources Blue/Green LED transilluminatorWhite LED transilluminatorWhite LED room light

Transilluminated area 26 x 21 cm (both)Fluorescent light 470-520 nmBlue/Green LEDs 24 x spots

Rated Voltage 100-240 V, 50 / 60 Hz, 2 A

Footprint (WxDxH) 38.2 cm x 40 cm x 78.5 cmWeight 35 Kg

Specifi cation

FAS-V

FAS-Digi

Key Features• Very high resolution - 16 MPixel LiveMOS Sensor

• 3” tiltable touchscreen display

• 20 cm x 16 cm Blue/Green LED Transilluminator

• White LED Transilluminator compatible

• Controlable by smartphones and tablets

The FastGene® FAS Digi was the first imaging system to use the Blue/Green LED imaging technology, and was designed to be a modular system. It is composed by the FastGene® Blue/Green LED Transilluminator (FG-08) and a high-end camera. It has the highest resolution of all of our imaging system with incredible 16 MPixel. The camera is Wi-Fi enabled and can be controlled using a smart device.

• Detection of red and green DNA/RNA dyes

• Detection of GFP, eGFP, YFP, etc.

• Documentation of protein gels

• Documentation of Petri dishes & membranes

• Excision of DNA from agarose gels

Applications

Image formats• Jpeg

• RAW

Blue/Green LED TransilluminatorUnique Excitation Technology

• Enables the detection of red and green DNA dyes

• Imaging area of 20 cm x 16 cm

• Superb uniform transillumination

• 6 large LED arrays at each side

Illumination

LiveMOS-Sensor• 24 mm - 64 mm focal length

(256 mm with digital zoom)

• 16 MPixel

• Focus by touchscreen

• Images can be recorded in JPEG and RAW format

Optical Unit

FAS Digi

16 MPixel LiveMOS Camera

Aluminum zoom ring

Amber fi lter

Metal dark box(removable)

Blue/Green LEDTransilluminator

3“ Touchscreen

Wi-Fi with tethering function

Highest resolution and ultra sharp pictures. Images can be printed in DIN A2 format

Ethidium Bromide MIDORIGreen Direct MIDORIGreen AdvanceEthidium Bromide MIDORIGreen Direct MIDORIGreen Advance

Comparison

B/G LED GelPicBox

Key Features• Smallest documentation system

• 2.7” LCD display

• 16 cm x 11.5 cm viewing area

• Blue/Green LED Transilluminator

• USB 2.0 port

The FastGene® Blue/Green LED GelPicBox combines a compact footprint with the advantages of the Blue/Green LED technology. This is the probably smallest imaging system in the world.The FastGene® Blue/Green LED GelPicBox is equipped with Blue/Green and white LEDs to document nucleic acids as well as protein gels and membranes.

• Detection of red and green DNA/RNA dyes

• Detection of GFP, eGFP, YFP, etc.

• Documentation of protein gels

• Documentation of Petri dishes & membranes

• Excision of DNA from agarose gels

Applications

Image formats• Jpeg

• TIFF

• BMP

Blue/Green LED TransilluminatorUnique Excitation Technology

• Enables the detection of red and green DNA dyes

• Imaging area of 16 cm x 11 cm

• Superb uniform transillumination

• 12 LED arrays at each side

White LED Lid• Detection of protein

• White LED in the lid

White LED Room light• Documentation of membranes and Petri dishes

• Two white LED arrays illuminate the imaging box

Illumination

CMOS-Sensor• Fixed focal length

• 9 MPixel

• Images can be recorded in TIFF, JPEG and BMP format

• External USB-stick

Optical Unit

Blue/Green & White light selection switch

Blue/Green & White light selection switch

B/G LED GelPicBox

Upper cover lid

White backlight

LED white light

Power switch

Scan button

Menu button

USB slot

LCD display

Ethidium Bromide MIDORIGreen Direct MIDORIGreen AdvanceEthidium Bromide MIDORIGreen Direct MIDORIGreen Advance

Comparison

FAS-Nano

Key Features• Very compact footprint

• Use any phone tablet with a camera.

• 10 cm x 10 cm Blue/Green LED Illuminator

• Any DNA dye

• Many different fluorescent protein

The FastGene® FAS-Nano is our smallest gel documentation system. It can be used as an illuminator or can be attached to a smartphone or tablet and be used as a gel documantation system.Filters and adapters are included.

• Detection and documentation of DNA/RNA dyes

• Detection of GFP, eGFP, YFP, etc.

• Documentation of fluorescent colonies on Petri

dishes & membranes

Applications

Blue/Green LED IlluminatorUnique Excitation Technology

• Enables the detection of red and green DNA dyes• Imaging area for gels of 10 x 10 cm gels• Superb illumination• Bright LEDs at each side

Lens• Very easy to mount on a smartphone or tablet• Comes with a holder for your smart device

Long pass filtersTwo different filter:

1) Amber filter shield: Large amber filter with long pass properties allowing light from 540 nm to pass. It is optimal for excising DNA bands out of the gel.

2) Amber lens filter: Extreme light passage properties for optimal detection and recording of images using a smartphone. The long pass properties allow light from 540 nm to pass.

Optical Unit

Illumination

Comparison

FAS-Nano has all the accessories needed to transform your smartphone into a gel documentation system.

Smart device holder

Lens

Filter holder

Amber Filter

Dark box

Blue/Green LED Illuminator

The Blue/Green LED technology is able to excite red and green DNA dyes, e.g. MIDORIGreen, SYBR™ & EtBr

FAS-Nano

You can use your normal photo app to take pictures of your gels.

MIDORIGreen Direct Ethidium Bromide

NGFG08 FG-11

NGFG05 NGFG06 FG-300

www.bulldog-bio.cominfo@bulldog-bio.com1-603-570-42481-603-766-0524

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