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Contents

Contents

1 Agilent DNA 7500 and DNA 12000 Kit

4

2 Equipment Required for a DNA 7500 and DNA 12000 Assay

6

3 Setting up the Assay Equipment and Bioanalyzer

7

Setting up the Chip Priming Station

8

Setting up the Bioanalyzer

9

Vortex Mixer

9

Starting the 2100 Expert Software

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 Samples

16

Inserting a Chip in the Agilent 2100 Bioanalyzer

17

Starting the Chip Run

18

Cleaning Electrodes after a Chip Run

20

6 Checking Your Agilent DNA 7500 and DNA 12000 Assay Results

21

DNA 7500 and DNA 12000 Ladder Well Results

21

DNA 7500 and DNA 12000 Sample Well Results

23

Содержание DNA 12000

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

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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...

Страница 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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