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Checking Your Agilent DNA 7500 and DNA 12000 Assay Results

DNA 7500 and DNA 12000 Ladder Well Results

Figure 3

DNA 12000 ladder

Major features of a successful ladder run are:

•

12 peaks for the DNA 7500 ladder and 13 peaks for the DNA 12000 ladder

•

All peaks are well resolved

•

Flat baseline

•

Correct identification of both markers

If the electropherogram of the ladder well window does not resemble the
one shown above, refer to the

2100 Expert Maintenance and

Troubleshooting Guide

for assistance.

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Summary of Contents for DNA 12000

Page 1: ...Agilent Technologies Agilent DNA 7500 and DNA 12000 Kit Guide...

Page 2: ...nd may be used or copied only in accordance with the terms of such license Restricted Rights Legend If software is for use in the performance of a U S Government prime contract or sub contract Softwar...

Page 3: ...e 10 4 Essential Measurement Practices 11 5 Agilent DNA 7500 and DNA 12000 Assay Protocol 12 Preparing the Gel Dye Mix 12 Loading the Gel Dye Mix 14 Loading the Marker 15 Loading the Ladder and the Sa...

Page 4: ...of this product conveys to the buyer the non transferable right to use the purchased amount of the product and components of the product only as described in accompanying product literature The sale o...

Page 5: ...DNA 12000 Sizing range 100 7500 bp 100 12000 bp Sizing resolution 100 1000 bp 5 1000 7500 bp 15 100 1000 bp 5 100 12000 bp 15 Sizing accuracy 10 15 Sizing reproducability 5 CV 5 CV Quantitation accur...

Page 6: ...g station reorder number 5065 4401 IKA vortex mixer Additional Material Required Not Supplied Pipettes 10 l 100 l and 1000 l with compatible tips Microcentrifuge tubes 0 5 ml for sample preparation 1...

Page 7: ...ace the syringe at the chip priming station with each new kit adjust the base plate of the chip priming station adjust the syringe clip at the chip priming station set up the vortex mixer finally make...

Page 8: ...new syringe and insert it into the clip d Slide it into the hole of the luer lock adapter and screw it tightly to the chip priming station 2 Adjust the base plate a Open the chip priming station by pu...

Page 9: ...yzer and make sure that the electrode cartridge is inserted in the instrument If not open the latch and insert the electrode cartridge Figure 1 Electrode cartridge inserted in the instrument graphic s...

Page 10: ...con The screen of the software appears in the Instrument context The icon in the upper part of the screen represents the current instrument PC communication status 2 If more than one instrument is con...

Page 11: ...ate to room temperature for 30 min before use Protect dye and dye mixtures from light Remove light covers only when pipetting The dye decomposes when exposed to light and this reduces the signal inten...

Page 12: ...brate to room temperature for 30 minutes WARNING Handling DMSO Kit components contain DMSO Because the dye binds to nucleic acids it should be treated as a potential mutagen and used with appropriate...

Page 13: ...e complete gel dye mix to the top receptacle of a spin filter 6 Place the spin filter in a microcentrifuge and spin for 10 minutes at room temperature at 1500 g 20 for Eppendorf microcentrifuge this c...

Page 14: ...ml and then close the chip priming station The lock of the latch will click when the chip priming station is closed correctly NOTE Before loading the gel dye mix make sure that the base plate of the c...

Page 15: ...to the 1 ml position 9 Open the chip priming station 10 Pipette 9 0 l of the gel dye mix in each of the wells marked Loading the Marker 1 Pipette 5 l of green capped marker green into the well marked...

Page 16: ...ixer and make sure not to damage the buldge that fixes the chip during vortexing 5 Vortex for 60 seconds at 2400 rpm 6 Refer to the next topic on how to insert the chip in the Agilent 2100 Bioanalyzer...

Page 17: ...e chip carefully into the receptacle The chip fits only one way 4 Carefully close the lid The electrodes in the cartridge fit into the wells of the chip 5 The 2100 expert software screen shows that yo...

Page 18: ...ix you have just entered At this time you can also customize the file storage location and the number of samples that will be analyzed NOTE Please note that the order of executing the chip run may cha...

Page 19: ...art the chip run The incoming raw signals are displayed in the Instrument context 5 After the chip run is finished remove the chip from the receptacle of the bioanalyzer and dispose of it according to...

Page 20: ...ectrode cleaner in the Agilent 2100 Bioanalyzer 3 Close the lid and leave it closed for about 10 seconds 4 Open the lid and remove the electrode cleaner 5 Wait another 10 seconds to allow the water on...

Page 21: ...Results 6 DNA 7500 and DNA 12000 Ladder Well Results To check the results of your run select the Gel or Electropherogram tab in the Data context The electropherogram of the ladder well window should r...

Page 22: ...ul ladder run are 12 peaks for the DNA 7500 ladder and 13 peaks for the DNA 12000 ladder All peaks are well resolved Flat baseline Correct identification of both markers If the electropherogram of the...

Page 23: ...features for a successful DNA sample run are All sample peaks appear between the lower and upper marker peaks If some sample peaks are outside the marker bracket adjust the upper or lower marker Plea...

Page 24: ...n Germany 12 2016 G2938 90024 G2938 90024 G2938 90024 www agilent com Agilent Technologies In This Book you find the procedures to analyze DNA samples with the Agilent DNA 7500 and DNA 12000 reagent k...

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