Name: simple and rapid method to obtain high-quality tumor dna from clinical-pathological specimens using touch imprint cytology
Uploaded: 2018-03-21T13:00:00
Description: Watch this Scientific Journal Video about Simple and Rapid Method to Obtain High-quality Tumor DNA from Clinical-pathological Specimens Using Touch Imprint Cytology at JoVE.com

Agradecemos a todo el personal médico y auxiliar del hospital y los pacientes para consentir a participar. Agradecemos a Gabrielle blanco lobo, PhD, de Edanz Group (www.edanzediting.com/ac) para la edición de un proyecto de este informe. Este estudio fue apoyado por una subvenciones para proyecto de investigación del genoma de la Prefectura de Yamanashi (Y.H. y M.O.) y una beca de la YASUDA médica Fundación (albergue).

1. TIC preparación para evaluación microscópica rápida utilizando vidrio Normal se desliza
<ol><li>Realizar la preparación de TIC tan pronto como sea posible después de tejido patológico clínico materiales están disponibles. Si muestras TIC no pueden estar preparadas inmediatamente, conserve los materiales de tejido cubiertos con solución salina humedecida gasa estéril y guarde en el refrigerador para prevenir la resequedad de los tejidos.</li>
	<li>Preparar 5 mm3 tejido material como tumores sól...

<ol>
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En este estudio, se presenta un método alternativo para la obtención de tumor ADN de especímenes patológicos clínicos utilizando TIC. Preparación de TIC es muy simple y necesita menos tiempo en comparación con métodos FFPE, sin la necesidad de instrumentos especiales10. Todos los procedimientos de la preparación de TIC para la extracción de ADN se pueden terminar dentro de dos días (figura 1). Este método por lo tanto acorta el tiempo de respuesta para rea...

Última generación tecnología de secuenciación ha proporcionado los investigadores avances significativos en el análisis de información del genoma en las variaciones genéticas, enfermedad mendeliana, predisposición hereditaria y cáncer 1,2,3 . El Atlas de genoma del cáncer (TCGA) y el Consorcio Internacional del genoma del cáncer (ICGC) han seguido la identificación de alteraciones genéticas en algunos tipos de cánceres comunes4. Cientos de genes de la conductor de cáncer esenciales han sido correctamente identificados y algunas de estas moléculas se apunta para drogas desarrollo1,5,6.
En el ajuste clínico, muestras FFPE comúnmente se utilizan para diagnosis patológica y moleculares para varias enfermedades, incluyendo cáncer. Sin embargo, durante el proceso de fijación con formalina, ADN-proteína o DNA-DNA Cross-linking ocurre y se induce la fragmentación del ADN. Así, muestras de ADN de FFPE no son siempre adecuadas para análisis genético debido a baja calidad y cantidad de ADN7,8,9. Además, tarda varios días para preparar a las muestras FFPE y habilidad técnica es necesario preparar correctamente las secciones. Por lo tanto, es conveniente desarrollar un método simple y rápido para obtener ADN intacto de alta calidad.
La citología es un método alternativo para el diagnóstico patológico. Preparación de la muestra citológica es un más simple, menos costoso y más rápido acercamiento comparado con FFPE preparación10. La técnica TIC se ha realizado sobre los nodos de linfa del centinela y los tejidos marginales de pacientes de cáncer de mama para el diagnóstico rápido intraoperatorio para algunos años11,12. Sin embargo, hay algunos informes que han examinado si el ADN genómico de alta calidad puede extraerse muestras TIC y utilizado para el posterior análisis genético. Las muestras citológicas son comúnmente teñidas con Papanicolaou (Pap) o tinción de Giemsa, y divulgado previamente que la cantidad y calidad del ADN extraído de muestras TIC (especialmente Giemsa-manchada las muestras) son superiores a las muestras obtenidas de FFPE los tejidos13. En comparación con la tinción de Papanicolau, tinción de Giemsa tiene una ventaja que requieren menos procedimientos de tinción. En la tinción de Pap, después las muestras se han fijado y teñido, debe ser montados con montaje medio (e.g., Malinol) para distinguir el contenido de la muestra, tales como las células tumorales, las células normales y células inflamatorias bajo un microscopio. Si la muestra de Pap está preparada sin el paso de montaje, es casi imposible observar las células bajo un microscopio porque la muestra se seca. En comparación, la coloración de Giemsa se observan en el estado seco, por lo tanto, el paso de montaje no es necesario para la rápida evaluación celular. Para microdisección, tinción de Giemsa es más conveniente porque requiere a especímenes secos.
En este informe, introducir un método simple y rápido para la preparación de especímenes TIC con la tinción de Giemsa y demuestran que el TIC es una mejor fuente de ADN en comparación con especímenes FFPE.

<table border="0" cellpadding="0" cellspacing="0" width="796">
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		<tr height="41">
			<td height="41" style="height:41px;width:283px;">FINE FROST white20 micro slide glass</td>
			<td style="width:191px;">Matsunami Glass ind, Ltd</td>
			<td style="width:143px;">SFF-011</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">Arcturus PEN Membrane Glass Slides&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td>LCM0522</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">Cyto Quick A solution</td>
			<td>Muto Pure Chemicals</td>
			<td>20571</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">Cyto Quick B solution</td>
			<td>Muto Pure Chemicals</td>
			<td>20581</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">May-Grunwald Solution</td>
			<td>Muto Pure Chemicals</td>
			<td>15053</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">Giemsa solution</td>
			<td>Muto Pure Chemicals</td>
			<td>15002</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">QIAamp DNA FFPE tissue kit</td>
			<td>Qiagen</td>
			<td>56404</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">TaqMan Fast Advanced Master Mix</td>
			<td>Thermo Fisher Scientific</td>
			<td>4444557</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">TaqMan RNase P Detection Reagents Kit&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td>4316831</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">TaqMan Assay from FFPE DNA QC Assay v2</td>
			<td>Thermo Fisher Scientific</td>
			<td>4324034</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">MicroAmp Fast Optical 96-Well Reaction Plate&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td>4346907</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">MicroAmp optical Adhesive Film</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td style="width:143px;">4311971</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">MicroMixer E36</td>
			<td style="width:191px;">TITEC</td>
			<td style="width:143px;">0027765-000</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">ViiA 7 Real-Time PCR System</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td style="width:143px;">VIIA7-03</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Himac CF16RXII</td>
			<td style="width:191px;">Hitachi-koki</td>
			<td style="width:143px;">CF16RII</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Ion Library TaqMan Quantitation Kit</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td style="width:143px;">4468802</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Ion AmpliSeq Cancer Hotspot Panel v2</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td style="width:143px;">4475346</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Ion AmpliSeq Library Kit 2.0</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td style="width:143px;">4480442</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Ion Xpress Barcode Adapters 1-16 Kit</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td>4471250</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Ion PGM Hi-Q View Sequencing Kit (200 base)</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td style="width:143px;">A30044</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Ion Chef System</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td>4484177</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Veriti 96-well Thermal Cycler</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td>Veriti200</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Ion 318 Chip Kit v2 BC</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td>4488150</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Ion PGM System</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td>PGM11-001</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Ion PGM Wash 2 Bottle kit</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td>A25591</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Agencourt&#8482; AMPure&#8482; XP Kit</td>
			<td style="width:191px;">Beckman Coulter</td>
			<td style="width:143px;">A63881</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">16-position Magnetic Stand</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td style="width:143px;">4457858</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Nonstick, RNase-free Microfuge Tubes, 1.5 mL (Low binding tube)</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td style="width:143px;">AM12450</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Nuclease-free water</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td style="width:143px;">AM9938</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">MicroAmp&#8482; Optical 96-well Reaction Plates</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td style="width:143px;">4306737</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">MicroAmp&#8482; Clear Adhesive Film</td>
			<td style="width:191px;">Thermo Fisher Scientific</td>
			<td style="width:143px;">4306311</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Agencourt&#8482; AMPure&#8482; XP Kit</td>
			<td style="width:191px;">Beckman Coulter</td>
			<td style="width:143px;">A63881</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Ethanol(99.5)</td>
			<td style="width:191px;">Nacalai Tesque</td>
			<td style="width:143px;">08948-25</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Sodium hydroxide (10M)</td>
			<td style="width:191px;">Sigma</td>
			<td style="width:143px;">72068</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">DTU-Neo</td>
			<td style="width:191px;">TAITEC</td>
			<td style="width:143px;">0063286-000</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">E-36&#160;</td>
			<td style="width:191px;">TAITEC</td>
			<td>0027765-000</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">ECLIPSE Ci-L</td>
			<td style="width:191px;">Nikon</td>
			<td style="width:143px;">704354</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">Pipet-Lite LTS Pipette L-2XLS+</td>
			<td>METTLER TOLEDO</td>
			<td>17014393</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">Pipet-Lite LTS Pipette L-10XLS+</td>
			<td>METTLER TOLEDO</td>
			<td>17014388</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">Pipet-Lite LTS Pipette L-20XLS+</td>
			<td>METTLER TOLEDO</td>
			<td>17014392</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">Pipet-Lite LTS Pipette L-100XLS+</td>
			<td>METTLER TOLEDO</td>
			<td>17014384</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">Pipet-Lite LTS Pipette L-200XLS+</td>
			<td>METTLER TOLEDO</td>
			<td>17014391</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">Pipet-Lite LTS Pipette L-1000XLS+</td>
			<td>METTLER TOLEDO</td>
			<td>17014382</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">petit-change</td>
			<td>WAKEN</td>
			<td>MODEL8864</td>
			<td style="width:180px;">Mini centrifuge</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">petit-incubator</td>
			<td>WAKEN</td>
			<td>WKN-2290</td>
			<td style="width:180px;">Air incubator</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">SensiCare Powder-free Nitrile Exam Gloves</td>
			<td style="width:191px;">MEDLINE</td>
			<td>SEM486802</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Sterile gauze</td>
			<td style="width:191px;">Osaki</td>
			<td style="width:143px;">11138</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Refrigerator MediCool</td>
			<td style="width:191px;">SANYO</td>
			<td style="width:143px;">MPR-312DCN-PJ</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">FEATHER TRIMMING BLAD</td>
			<td style="width:191px;">FEATHER</td>
			<td style="width:143px;">No.130</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">FEATHER TRIMMING BLAD</td>
			<td style="width:191px;">FEATHER</td>
			<td style="width:143px;">No.260</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">FEATHER&#160; S</td>
			<td style="width:191px;">FEATHER</td>
			<td style="width:143px;">FA-10</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Vortex Genius 3</td>
			<td style="width:191px;">IKA</td>
			<td>41-0458&#160;</td>
			<td style="width:180px;">Vortex mixer</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">Pincette</td>
			<td style="width:191px;">NATSUME</td>
			<td style="width:143px;">A-5</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:283px;">1.5 mL microtube</td>
			<td style="width:191px;">BIOBIK</td>
			<td style="width:143px;">RC-0150</td>
			<td style="width:180px;"></td>
		</tr>
	</tbody>
</table>

simple and rapid method to obtain high-quality tumor dna from clinical-pathological specimens using touch imprint cytology

Es fundamental para determinar el estado mutacional en cáncer antes de la administración y tratamiento de drogas específicas moleculares específicas para pacientes con cáncer. En el ajuste clínico, tejidos de (FFPE) parafina-encajado formalina-fijos son ampliamente utilizados para las pruebas genéticas. Sin embargo, FFPE ADN generalmente dañado y fragmentado durante el proceso de fijación con formalina. Por lo tanto, FFPE ADN a veces no es adecuado para las pruebas genéticas debido a baja calidad y cantidad de ADN. Aquí presentamos un método de la citología de impresión táctil (TIC) para obtener ADN genómico de las células cancerosas, que puede ser observado bajo el microscopio. Número de células de cáncer y morfología celular puede ser evaluado utilizando a muestras TIC. Además, la extracción de DNA genómico de muestras TIC se puede terminar dentro de dos días. La cantidad total y calidad de ADN TIC obtenidos mediante este método fue mayor que el de ADN FFPE. Este método rápido y simple permite a los investigadores para obtener ADN de calidad para pruebas genéticas (p. ej., análisis de secuenciación de próxima generación, digital PCR y PCR cuantitativo en tiempo real) y para acortar el tiempo de respuesta para informar los resultados.

La figura 1 muestra el proceso de preparación de muestras TIC para la extracción de ADN. En particular, el procedimiento toma sólo dos días para obtener ADN genómico de muestras TIC. Se evaluaron los efectos de almacenamiento de tumor antes de la elaboración de diapositivas. Encontramos que las células del tumor fueron atadas en el portaobjetos de cristal cuando muestras de tejido fueron inmediatamente tocadas en la diapositiva, y cuando los tejidos se...

Watch this Scientific Journal Video about Simple and Rapid Method to Obtain High-quality Tumor DNA from Clinical-pathological Specimens Using Touch Imprint Cytology at JoVE.com

Simple and Rapid Method to Obtain High-quality Tumor DNA from Clinical-pathological Specimens Using Touch Imprint Cytology

Obtención de ADN genómico de la alta calidad de los tejidos del tumor es un primer paso esencial para el análisis de alteraciones genéticas mediante la siguiente secuencia de generación. En este artículo presentamos un método simple y rápido para enriquecer las células tumorales y obtener ADN intacto del toque de muestras de citología de impresión.

Método simple y rápido para obtener alta calidad Tumor ADN de especímenes clínicos patológicos mediante citología de impresión táctil

It is critical to determine the mutational status in cancer before administration and treatment of specific molecular targeted drugs for cancer patients. ...

The overall goal of this protocol is to obtain high-quality tumor DNA from clinical samples. This method can help answer key questions in basic and clinical science such as cancer research, diagnosis, and treatment decisions. The main advantage of this technique is that we can obtain tumor DNA more simply and quickly than with previous methods.To begin the protocol, touch the tumor surface of a previously prepared and resected tissue onto a normal glass slide several times with gloved hands. Visually confirm that the touched area is covering more than 80%of the glass slide. Lightly press the normal glass slide against a Polyethylene Naphthalate or PEN membrane slide and rub it gently two to three times with gloved hands.Visually confirm that the cells are transferred from the normal glass slide to the PEN membrane slide. Air dry both the glass and PEN membrane slides for five minutes at room temperature. After drying, stain the normal glass slide for direct cytological examination.Dip the glass slides into fixative solution for five seconds and then stain with Giemsa staining solution for 15 seconds. Assess and screen the tumor contents and cellularity on the normal glass slide entirely with a microscope for quick assessment. Evaluate tumor cells based on several criteria.If the sample shows tumor cellularity over 60%by quick microscopic assessment from the previous section, cut the tumor-touched film of the PEN membrane slides for DNA extraction with a knife and gloved hands. Transfer the cut film to a sterile microcentrifuge tube with a pin set and gloved hands. Store the film-containing microcentrifuge tube at four degrees Celsius until DNA extraction.Open the software and plan the run condition and set the run parameter within the software. Click the plan tab and select template, then select the appropriate run method. Select the application and technique type and click next.Select the instrument, sample preparation kit, library kit type, template kit, sequencing kit, base calibration mode, chip type, control sequence, and barcode set and click next. Select plugins and click next. Select project and click next.Then select default reference and BED files of the targeted region. Type the sample name, select the barcode, and click plan run. Next, perform template preparation and chip loading in an automated instrument according to the manufacturer's instructions.Thaw the reagent cartridge at room temperature for 45 minutes before use. Dilute the undiluted library with nuclease-free water according to the library concentration calculated in step 6.8 and make 20 picomolar libraries. Prepare a pooled library for sequencing and store it on ice.Add 25 microliters of the pooled library with a 100 microliter pipette to the bottom of the sample tube. Turn on the power and open the cover of the automated instrument. Place the sequencing chip, chip adapter, enrichment cartridge, tip cartridge, PCR plate, PCR frame seal, recovery tube, solution cartridge, and reagent cartridge to the appropriate position of the automated instrument.Touch set up run and step by step on the screen. Close the cover and touch start check on the screen. After the deck scan process, touch next.Check the display contents, kit type, chip ID, sample ID, chip type, plans. Set the time and touch OK.After finishing the chip loading, touch next and open the cover. Unload the sequencing chip from the chip adapter with gloved hands.Place the chip into the chip container, wrap it with parafilm, and store the chip at four degrees Celsius until the sequencing reaction. Then remove the enrichment cartridge, PCR plate, PCR frame seal, recovery tube, solution cartridge, and reagent cartridge from the appropriate position of the automated instrument with gloved hands. Transfer an empty tip cartridge to the waste tip position of the automated instrument with gloved hands.Touch next and close the cover. Then touch start and clean the automated instrument with ultraviolet rays for four minutes. Dissolve a sodium chlorite tablet in 1, 000 milliliters of ultra-pure water and filter the solution with a 0.22 micron filter.Once the sequencing instrument has warmed up, touch clean and next on the screen of the sequencing instrument. Clean the sequencing instrument with 250 milliliters of filter-sterilized sodium chlorite solution, then 250 milliliters of ultra-pure water. Then touch initialize and select the appropriate sequencing kit on the screen.Install a gray shipper with gloved hands. Initialize the sequencing instrument with the wash solution, the pH adjustment solution, and the pH standard solution. Next, add 20 microliters of DATP, DGTP, DCTP, and DTTP nucleotides in 50 milliliter tubes with a 100 microliter pipette.Install a gray shipper at the appropriate location with gloved hands. Load the 50 milliliter tube and screw it onto the sequencing instrument. Select next on the screen to start the initialization step.After completing the initialization step, touch run on the screen and select the appropriate library preparation instrument. Scan the two-dimensional barcode of the chip. Insert the sequencing chip in the appropriate position.Close the chip clamp and instrument door and touch chip check, next, and OK on the screen to start the sequencing run. After the sequencing reaction, transfer the data and perform data analyzing pipeline on the sequencing server. The absolute DNA quantities and the RQ value, an indicator of the degradation level of genomic DNA, show that a higher DNA yield is achieved using the TIC specimens compared with the FFPE specimens.In addition, the RQ values from the TIC DNA were significantly higher compared with that of the FFPE DNA across different tumor types. APC Q1367*was identified in both FFPE and TIC DNA extracted from site one and APC S1356*KRAS G12D, and TP53 M237i were detected in both FFPE and TIC DNA extracted from site two, three, and four suggesting that the identical somatic mutations were identified in paired FFPE and TIC DNA samples prepared from the same tumor site. Notably, the same somatic mutations were detected between primary colorectal cancer and two metastatic liver cancer samples suggesting that the tumor clones from site two metastasized to liver.After its development, this technique paved the way for researchers in the field of molecular and medical research to identify genetic cancers.

Research

JoVE Journal

Cancer Research

A Mimic of the Tumor Microenvironment: A Simple Method for Generating Enriched Cell Populations and Investigating Intercellular Communication

Un imitador del microambiente tumoral: Un método simple para generar poblaciones de células enriquecido y Comunicación intercelular Investigando

Understanding the early heterotypic interactions between cancer cells and the surrounding non-cancerous stroma is important in elucidating the events ...

Investigación

Investigación sobre el cáncer

Characterization of Tumor Cells Using a Medical Wire for Capturing Circulating Tumor Cells: A 3D Approach Based on Immunofluorescence and DNA FISH

Caracterización de células del Tumor mediante un alambre de médico para la captura de circulación de las células del Tumor: un enfoque 3D basada en la inmunofluorescencia y DNA FISH

Circulating tumor cells (CTCs) are associated with poor survival in metastatic cancer. Their identification, phenotyping, and genotyping could lead to a ...

Using 22C3 Anti-PD-L1 Antibody Concentrate on Biopsy and Cytology Samples from Non-small Cell Lung Cancer Patients

Con 22 3 anticuerpos Anti-PD-L1 concentrarse en biopsia y muestras citológicas de pacientes con cáncer pulmonar de células no pequeñas

Pembrolizumab monotherapy has been approved for the first- and second-line treatment of patients with PD-L1-expressing advanced non-small cell lung cancer ...

Digital Polymerase Chain Reaction Assay for the Genetic Variation in a Sporadic Familial Adenomatous Polyposis Patient Using the Chip-in-a-tube Format

Digital de la reacción en cadena de polimerasa para la variación genética en un paciente de la poliposis adenomatosa familiar esporádico con el formato de Chip en tubo

The quantitative analysis of human genetic variation is crucial for understanding the molecular characteristics of serious medical conditions, such as ...

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Detección y seguimiento del ADN circulante asociado a tumores en biofluidos del paciente

Complications associated with upfront and repeat surgical tissue sampling present the need for minimally invasive platforms capable of molecular ...

Rapid Optical Clearing for Semi-High-Throughput Analysis of Tumor Spheroids

Limpieza óptica rápida para el análisis de semi-alto rendimiento de esferoides tumorales

Tumor spheroids are fast becoming commonplace in basic cancer research and drug development. Obtaining data regarding protein expression within the ...

Culture and Imaging of Ex Vivo Organotypic Pseudomyxoma Peritonei Tumor Slices from Resected Human Tumor Specimens

Culture and Imaging of Ex Vivo Organotypic Pseudomyxoma Peritonei Tumor Slices from Resected Human Tumor Specimens

Cultivo e imágenes de cortes tumorales organotípicos ex vivo de pseudomixoma peritonei a partir de muestras tumorales humanas resecadas

Pseudomyxoma peritonei (PMP) is a rare condition that results from the dissemination of a mucinous primary tumor and the resultant accumulation of ...

Cuantificación de la longitud de los telómeros mediante la detección de quimioluminiscencia no radiactiva

Optimization of Performance Parameters of the TAGGG Telomere Length Assay

Autor destacado: Optimización de los parámetros de rendimiento del ensayo de longitud de telómeros TAGGG  

Optimización de los parámetros de rendimiento del ensayo de longitud de telómeros TAGGG

Telomeres are repetitive sequences which are present at chromosomal ends; their shortening is a characteristic feature of human somatic cells. Shortening ...