Structured Review

Agilent technologies bioanalyzer 2100 instrument
( a ) Multiplex ligation-dependent genome amplification (MLGA), reaction scheme.  (I)  Genomic DNA is digested by restriction enzyme to generate targets with defined ends.  (II)  Each MLGA probe consists of two oligonucleotides, one selector oligo of 70–74 nt (green) and one general vector oligo of 34 nt (red). MLGA probe together with DNA-ligase forms circular DNA of target molecules after denaturation and hybridization.  (III)  To reduce background signal in the assay, undesirable, linear DNA is degraded by exonuclease I (Exo I).  (IV)  Multiplex PCR is facilitated by using universal primers that hybridize to a sequence in the vector. PCR products are analyzed using the Agilent Bioanalyzer 2100™ electrophoresis system. ( b ) Data from an MLGA set of 14 probes targeting loci on human chromosomes 13, 18, 21, X and Y. The upper graph shows the resulting elution diagrams from analyses of male and female DNA pools.
Bioanalyzer 2100 Instrument, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 94/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/bioanalyzer 2100 instrument/product/Agilent technologies
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Price from $9.99 to $1999.99
bioanalyzer 2100 instrument - by Bioz Stars, 2020-05
94/100 stars

Images

1) Product Images from "MLGA--a rapid and cost-efficient assay for gene copy-number analysis"

Article Title: MLGA--a rapid and cost-efficient assay for gene copy-number analysis

Journal: Nucleic Acids Research

doi: 10.1093/nar/gkm651

( a ) Multiplex ligation-dependent genome amplification (MLGA), reaction scheme.  (I)  Genomic DNA is digested by restriction enzyme to generate targets with defined ends.  (II)  Each MLGA probe consists of two oligonucleotides, one selector oligo of 70–74 nt (green) and one general vector oligo of 34 nt (red). MLGA probe together with DNA-ligase forms circular DNA of target molecules after denaturation and hybridization.  (III)  To reduce background signal in the assay, undesirable, linear DNA is degraded by exonuclease I (Exo I).  (IV)  Multiplex PCR is facilitated by using universal primers that hybridize to a sequence in the vector. PCR products are analyzed using the Agilent Bioanalyzer 2100™ electrophoresis system. ( b ) Data from an MLGA set of 14 probes targeting loci on human chromosomes 13, 18, 21, X and Y. The upper graph shows the resulting elution diagrams from analyses of male and female DNA pools.
Figure Legend Snippet: ( a ) Multiplex ligation-dependent genome amplification (MLGA), reaction scheme. (I) Genomic DNA is digested by restriction enzyme to generate targets with defined ends. (II) Each MLGA probe consists of two oligonucleotides, one selector oligo of 70–74 nt (green) and one general vector oligo of 34 nt (red). MLGA probe together with DNA-ligase forms circular DNA of target molecules after denaturation and hybridization. (III) To reduce background signal in the assay, undesirable, linear DNA is degraded by exonuclease I (Exo I). (IV) Multiplex PCR is facilitated by using universal primers that hybridize to a sequence in the vector. PCR products are analyzed using the Agilent Bioanalyzer 2100™ electrophoresis system. ( b ) Data from an MLGA set of 14 probes targeting loci on human chromosomes 13, 18, 21, X and Y. The upper graph shows the resulting elution diagrams from analyses of male and female DNA pools.

Techniques Used: Multiplex Assay, Ligation, Amplification, Plasmid Preparation, Hybridization, Polymerase Chain Reaction, Sequencing, Electrophoresis

2) Product Images from "Mixed Triple: Allied Viruses in Unique Recent Isolates of Highly Virulent Type 2 Bovine Viral Diarrhea Virus Detected by Deep Sequencing"

Article Title: Mixed Triple: Allied Viruses in Unique Recent Isolates of Highly Virulent Type 2 Bovine Viral Diarrhea Virus Detected by Deep Sequencing

Journal: Journal of Virology

doi: 10.1128/JVI.00620-14

Agilent Bioanalyzer 2100 analyses (with a DNA 7500 chip) of PCR products. (Upper panels) Gel view of Agilent traces. Bands representing dup + and dup − genomes are marked by arrows labeled accordingly. (Lower panels) Bar plots of relative quantities
Figure Legend Snippet: Agilent Bioanalyzer 2100 analyses (with a DNA 7500 chip) of PCR products. (Upper panels) Gel view of Agilent traces. Bands representing dup + and dup − genomes are marked by arrows labeled accordingly. (Lower panels) Bar plots of relative quantities

Techniques Used: Chromatin Immunoprecipitation, Polymerase Chain Reaction, Labeling

3) Product Images from "Human Milk MicroRNA and Total RNA Differ Depending on Milk Fractionation"

Article Title: Human Milk MicroRNA and Total RNA Differ Depending on Milk Fractionation

Journal: Journal of Cellular Biochemistry

doi: 10.1002/jcb.25207

Comparison of overall concentration of total RNA using NanoDrop 1000, and microRNA using Bioanalyzer 2100, respectively, between the lipid and skim milk fractions obtained for all samples using the eight extraction kits.
Figure Legend Snippet: Comparison of overall concentration of total RNA using NanoDrop 1000, and microRNA using Bioanalyzer 2100, respectively, between the lipid and skim milk fractions obtained for all samples using the eight extraction kits.

Techniques Used: Concentration Assay

4) Product Images from "A novel approach to stabilize fetal cell-free DNA fraction in maternal blood samples for extended period of time"

Article Title: A novel approach to stabilize fetal cell-free DNA fraction in maternal blood samples for extended period of time

Journal: PLoS ONE

doi: 10.1371/journal.pone.0208508

Analysis of plasma cfDNA obtained from one representative pregnant donor blood stored at 22°C using Agilent Bioanalyzer 2100 instrument and Agilent DNA high sensitivity Kit. A, Overlaid electropherograms of plasma cfDNA extracted from blood stored (at 22°C) in K 3 EDTA tubes at days 0, 3, 7, 14 and 28. B, Overlaid electropherograms of plasma cfDNA extracted from blood stored (at 22°C) in ProTeck tubes at days 0, 3, 7, 14 and 28. C, Bioanalyzer gel image for blood stored in K 3 EDTA tubes. D, Bioanalyzer gel image for blood stored in ProTeck tubes. Cell-free DNA obtained from 3 pregnant donors were analyzed. This figure shows only the results of one representative pregnant donor.
Figure Legend Snippet: Analysis of plasma cfDNA obtained from one representative pregnant donor blood stored at 22°C using Agilent Bioanalyzer 2100 instrument and Agilent DNA high sensitivity Kit. A, Overlaid electropherograms of plasma cfDNA extracted from blood stored (at 22°C) in K 3 EDTA tubes at days 0, 3, 7, 14 and 28. B, Overlaid electropherograms of plasma cfDNA extracted from blood stored (at 22°C) in ProTeck tubes at days 0, 3, 7, 14 and 28. C, Bioanalyzer gel image for blood stored in K 3 EDTA tubes. D, Bioanalyzer gel image for blood stored in ProTeck tubes. Cell-free DNA obtained from 3 pregnant donors were analyzed. This figure shows only the results of one representative pregnant donor.

Techniques Used:

Analysis of plasma cfDNA obtained from one representative pregnant donor blood stored at 30°C using Agilent Bioanalyzer 2100 instrument and Agilent DNA high sensitivity Kit. A, Overlaid electropherograms of plasma cfDNA extracted from blood stored in K 3 EDTA tubes at days 0, 2, 3, 7 and 14. B, Overlaid electropherograms of plasma cfDNA extracted from blood stored in ProTeck tubes at days 0, 2, 3, 7 and 14. C, Bioanalyzer gel image for blood stored in K 3 EDTA tubes. D, Bioanalyzer gel image for blood stored in ProTeck tubes. Cell-free DNA obtained from 3 pregnant donors were analyzed. This figure shows only the results of one representative pregnant donor.
Figure Legend Snippet: Analysis of plasma cfDNA obtained from one representative pregnant donor blood stored at 30°C using Agilent Bioanalyzer 2100 instrument and Agilent DNA high sensitivity Kit. A, Overlaid electropherograms of plasma cfDNA extracted from blood stored in K 3 EDTA tubes at days 0, 2, 3, 7 and 14. B, Overlaid electropherograms of plasma cfDNA extracted from blood stored in ProTeck tubes at days 0, 2, 3, 7 and 14. C, Bioanalyzer gel image for blood stored in K 3 EDTA tubes. D, Bioanalyzer gel image for blood stored in ProTeck tubes. Cell-free DNA obtained from 3 pregnant donors were analyzed. This figure shows only the results of one representative pregnant donor.

Techniques Used:

Analysis of plasma cfDNA obtained from one representative pregnant donor blood stored at 4°C using Agilent Bioanalyzer 2100 instrument and Agilent DNA high sensitivity Kit. A, Overlaid electropherograms of plasma cfDNA extracted from blood stored in K 3 EDTA tubes at days 0, 3, 7, 14 and 21. B, Overlaid electropherograms of plasma cfDNA extracted from blood stored in ProTeck tubes at days 0, 3, 7, 14 and 21. C, Bioanalyzer gel image for blood stored in K 3 EDTA tubes. D, Bioanalyzer gel image for blood stored in ProTeck tubes. Cell-free DNA obtained from 3 pregnant donors were analyzed. This figure shows only the results of one representative pregnant donor.
Figure Legend Snippet: Analysis of plasma cfDNA obtained from one representative pregnant donor blood stored at 4°C using Agilent Bioanalyzer 2100 instrument and Agilent DNA high sensitivity Kit. A, Overlaid electropherograms of plasma cfDNA extracted from blood stored in K 3 EDTA tubes at days 0, 3, 7, 14 and 21. B, Overlaid electropherograms of plasma cfDNA extracted from blood stored in ProTeck tubes at days 0, 3, 7, 14 and 21. C, Bioanalyzer gel image for blood stored in K 3 EDTA tubes. D, Bioanalyzer gel image for blood stored in ProTeck tubes. Cell-free DNA obtained from 3 pregnant donors were analyzed. This figure shows only the results of one representative pregnant donor.

Techniques Used:

5) Product Images from "A novel approach to stabilize fetal cell-free DNA fraction in maternal blood samples for extended period of time"

Article Title: A novel approach to stabilize fetal cell-free DNA fraction in maternal blood samples for extended period of time

Journal: PLoS ONE

doi: 10.1371/journal.pone.0208508

Analysis of plasma cfDNA obtained from one representative pregnant donor blood stored at 22°C using Agilent Bioanalyzer 2100 instrument and Agilent DNA high sensitivity Kit. A, Overlaid electropherograms of plasma cfDNA extracted from blood stored (at 22°C) in K 3 EDTA tubes at days 0, 3, 7, 14 and 28. B, Overlaid electropherograms of plasma cfDNA extracted from blood stored (at 22°C) in ProTeck tubes at days 0, 3, 7, 14 and 28. C, Bioanalyzer gel image for blood stored in K 3 EDTA tubes. D, Bioanalyzer gel image for blood stored in ProTeck tubes. Cell-free DNA obtained from 3 pregnant donors were analyzed. This figure shows only the results of one representative pregnant donor.
Figure Legend Snippet: Analysis of plasma cfDNA obtained from one representative pregnant donor blood stored at 22°C using Agilent Bioanalyzer 2100 instrument and Agilent DNA high sensitivity Kit. A, Overlaid electropherograms of plasma cfDNA extracted from blood stored (at 22°C) in K 3 EDTA tubes at days 0, 3, 7, 14 and 28. B, Overlaid electropherograms of plasma cfDNA extracted from blood stored (at 22°C) in ProTeck tubes at days 0, 3, 7, 14 and 28. C, Bioanalyzer gel image for blood stored in K 3 EDTA tubes. D, Bioanalyzer gel image for blood stored in ProTeck tubes. Cell-free DNA obtained from 3 pregnant donors were analyzed. This figure shows only the results of one representative pregnant donor.

Techniques Used:

Analysis of plasma cfDNA obtained from one representative pregnant donor blood stored at 30°C using Agilent Bioanalyzer 2100 instrument and Agilent DNA high sensitivity Kit. A, Overlaid electropherograms of plasma cfDNA extracted from blood stored in K 3 EDTA tubes at days 0, 2, 3, 7 and 14. B, Overlaid electropherograms of plasma cfDNA extracted from blood stored in ProTeck tubes at days 0, 2, 3, 7 and 14. C, Bioanalyzer gel image for blood stored in K 3 EDTA tubes. D, Bioanalyzer gel image for blood stored in ProTeck tubes. Cell-free DNA obtained from 3 pregnant donors were analyzed. This figure shows only the results of one representative pregnant donor.
Figure Legend Snippet: Analysis of plasma cfDNA obtained from one representative pregnant donor blood stored at 30°C using Agilent Bioanalyzer 2100 instrument and Agilent DNA high sensitivity Kit. A, Overlaid electropherograms of plasma cfDNA extracted from blood stored in K 3 EDTA tubes at days 0, 2, 3, 7 and 14. B, Overlaid electropherograms of plasma cfDNA extracted from blood stored in ProTeck tubes at days 0, 2, 3, 7 and 14. C, Bioanalyzer gel image for blood stored in K 3 EDTA tubes. D, Bioanalyzer gel image for blood stored in ProTeck tubes. Cell-free DNA obtained from 3 pregnant donors were analyzed. This figure shows only the results of one representative pregnant donor.

Techniques Used:

Analysis of plasma cfDNA obtained from one representative pregnant donor blood stored at 4°C using Agilent Bioanalyzer 2100 instrument and Agilent DNA high sensitivity Kit. A, Overlaid electropherograms of plasma cfDNA extracted from blood stored in K 3 EDTA tubes at days 0, 3, 7, 14 and 21. B, Overlaid electropherograms of plasma cfDNA extracted from blood stored in ProTeck tubes at days 0, 3, 7, 14 and 21. C, Bioanalyzer gel image for blood stored in K 3 EDTA tubes. D, Bioanalyzer gel image for blood stored in ProTeck tubes. Cell-free DNA obtained from 3 pregnant donors were analyzed. This figure shows only the results of one representative pregnant donor.
Figure Legend Snippet: Analysis of plasma cfDNA obtained from one representative pregnant donor blood stored at 4°C using Agilent Bioanalyzer 2100 instrument and Agilent DNA high sensitivity Kit. A, Overlaid electropherograms of plasma cfDNA extracted from blood stored in K 3 EDTA tubes at days 0, 3, 7, 14 and 21. B, Overlaid electropherograms of plasma cfDNA extracted from blood stored in ProTeck tubes at days 0, 3, 7, 14 and 21. C, Bioanalyzer gel image for blood stored in K 3 EDTA tubes. D, Bioanalyzer gel image for blood stored in ProTeck tubes. Cell-free DNA obtained from 3 pregnant donors were analyzed. This figure shows only the results of one representative pregnant donor.

Techniques Used:

Related Articles

Electrophoresis:

Article Title: APOBEC3A cytidine deaminase induces RNA editing in monocytes and macrophages
Article Snippet: .. Electrophoresis of the libraries on Bioanalyzer 2100 instrument (Agilent, Santa Clara, CA) showed highest peaks at 220–240 bp. .. Paired-end multiplexed sequencing of libraries (three per flow cell lane) to generate reads of 101 b was performed on HiSeq 2000 instrument with TruSeq SBS and PE Cluster v3 Kits (Illumina).

Article Title: Human Milk MicroRNA and Total RNA Differ Depending on Milk Fractionation
Article Snippet: .. The microRNA concentration and microRNA/small RNA ratios were quantified by capillary electrophoresis using the small RNA Chip kit (Agilent, CA) in an Agilent Bioanalyzer 2100 instrument. ..

Concentration Assay:

Article Title: Epigenetic Modification of the Glucocorticoid Receptor Gene Is Linked to Traumatic Memory and Post-Traumatic Stress Disorder Risk in Genocide Survivors
Article Snippet: .. Bisulfite DNA quality and concentration was determined using an RNA Pico 6000 Kit on a Bioanalyzer 2100 instrument (Agilent Technologies) and Nanodrop 2000 (ThermoScientific). ..

Article Title: Human Milk MicroRNA and Total RNA Differ Depending on Milk Fractionation
Article Snippet: .. The microRNA concentration and microRNA/small RNA ratios were quantified by capillary electrophoresis using the small RNA Chip kit (Agilent, CA) in an Agilent Bioanalyzer 2100 instrument. ..

Polymerase Chain Reaction:

Article Title: Mixed Triple: Allied Viruses in Unique Recent Isolates of Highly Virulent Type 2 Bovine Viral Diarrhea Virus Detected by Deep Sequencing
Article Snippet: .. PCR products were analyzed with a Bioanalyzer 2100 instrument (Agilent Technologies, Böblingen, Germany), using an Agilent DNA 7500 chip. .. Quantification of the PCR products was done using 2100 Expert software (version B.02.08.SI648 [SR2]; Agilent).

Article Title: MLGA--a rapid and cost-efficient assay for gene copy-number analysis
Article Snippet: .. Temperature cycling was performed as follows: 37° C for 30 min, 95° C for 5 min followed by 30 cycles of 95° C 15 s, 55° C 30 s and 72° C for 60 s followed by 72° C for 10 min. PCR products were analyzed using an Agilent Bioanalyzer 2100™ instrument and quantified using the Agilent 2100 expert software, version B.02.02.SI238. ..

Chromatin Immunoprecipitation:

Article Title: Human Milk MicroRNA and Total RNA Differ Depending on Milk Fractionation
Article Snippet: .. The microRNA concentration and microRNA/small RNA ratios were quantified by capillary electrophoresis using the small RNA Chip kit (Agilent, CA) in an Agilent Bioanalyzer 2100 instrument. ..

Article Title: Mixed Triple: Allied Viruses in Unique Recent Isolates of Highly Virulent Type 2 Bovine Viral Diarrhea Virus Detected by Deep Sequencing
Article Snippet: .. PCR products were analyzed with a Bioanalyzer 2100 instrument (Agilent Technologies, Böblingen, Germany), using an Agilent DNA 7500 chip. .. Quantification of the PCR products was done using 2100 Expert software (version B.02.08.SI648 [SR2]; Agilent).

Software:

Article Title: MLGA--a rapid and cost-efficient assay for gene copy-number analysis
Article Snippet: .. Temperature cycling was performed as follows: 37° C for 30 min, 95° C for 5 min followed by 30 cycles of 95° C 15 s, 55° C 30 s and 72° C for 60 s followed by 72° C for 10 min. PCR products were analyzed using an Agilent Bioanalyzer 2100™ instrument and quantified using the Agilent 2100 expert software, version B.02.02.SI238. ..

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    Agilent technologies 2100 bioanalyzer instrument
    Mutation g.chrX:70339329T > C in MED12 causes the splicing of exon 2 ( A ) Schematic representation of the normal (top) and aberrant splicing (bottom) of MED12 transcripts from the normal and the identified mutant gene. In the presence of the mutation, the donor splice site of exon 2 (underlined) is skipped and exon 1 is directly spliced to exon 3, leading to an aberrant mature form of MED12 mRNA lacking exon 2. ( B ) Synthetic electrophoresis gel of MED12 PCR products (Methods). cDNAs were synthesized from mature RNA extracted from patient 744 mutated at g.chrX :70339329T > C, REH cells, mature T-cells (CD19-CD3+) isolated from cord blood samples and two T-ALL patients, WT for this position (791 and 727). Amplified fragments of 281 bp (WT samples) and 176 bp (mutated sample) were analyzed using Agilent <t>2100</t> <t>Bioanalyzer.</t> The 105 bp difference corresponds to the skipped exon 2. L: ladder; CTL-: negative control. ( C ) Screenshot of the Integrative Genomics Viewer (IGV) window presenting aligned RNA-seq data obtained from cases 744 (top) and a third T-ALL WT patient (693, bottom). The dotted line is centered on the donor splice site of exon 2 of MED12 (g.chrX:70339329). Case 744 shows aberrant splicing of exon 2, while 693 was WT for this position and shows a mature transcript that includes the exon 2.
    2100 Bioanalyzer Instrument, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 94/100, based on 92 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/2100 bioanalyzer instrument/product/Agilent technologies
    Average 94 stars, based on 92 article reviews
    Price from $9.99 to $1999.99
    2100 bioanalyzer instrument - by Bioz Stars, 2020-05
    94/100 stars
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    Mutation g.chrX:70339329T > C in MED12 causes the splicing of exon 2 ( A ) Schematic representation of the normal (top) and aberrant splicing (bottom) of MED12 transcripts from the normal and the identified mutant gene. In the presence of the mutation, the donor splice site of exon 2 (underlined) is skipped and exon 1 is directly spliced to exon 3, leading to an aberrant mature form of MED12 mRNA lacking exon 2. ( B ) Synthetic electrophoresis gel of MED12 PCR products (Methods). cDNAs were synthesized from mature RNA extracted from patient 744 mutated at g.chrX :70339329T > C, REH cells, mature T-cells (CD19-CD3+) isolated from cord blood samples and two T-ALL patients, WT for this position (791 and 727). Amplified fragments of 281 bp (WT samples) and 176 bp (mutated sample) were analyzed using Agilent 2100 Bioanalyzer. The 105 bp difference corresponds to the skipped exon 2. L: ladder; CTL-: negative control. ( C ) Screenshot of the Integrative Genomics Viewer (IGV) window presenting aligned RNA-seq data obtained from cases 744 (top) and a third T-ALL WT patient (693, bottom). The dotted line is centered on the donor splice site of exon 2 of MED12 (g.chrX:70339329). Case 744 shows aberrant splicing of exon 2, while 693 was WT for this position and shows a mature transcript that includes the exon 2.

    Journal: Oncotarget

    Article Title: Genomic characterization of pediatric T-cell acute lymphoblastic leukemia reveals novel recurrent driver mutations

    doi: 10.18632/oncotarget.11796

    Figure Lengend Snippet: Mutation g.chrX:70339329T > C in MED12 causes the splicing of exon 2 ( A ) Schematic representation of the normal (top) and aberrant splicing (bottom) of MED12 transcripts from the normal and the identified mutant gene. In the presence of the mutation, the donor splice site of exon 2 (underlined) is skipped and exon 1 is directly spliced to exon 3, leading to an aberrant mature form of MED12 mRNA lacking exon 2. ( B ) Synthetic electrophoresis gel of MED12 PCR products (Methods). cDNAs were synthesized from mature RNA extracted from patient 744 mutated at g.chrX :70339329T > C, REH cells, mature T-cells (CD19-CD3+) isolated from cord blood samples and two T-ALL patients, WT for this position (791 and 727). Amplified fragments of 281 bp (WT samples) and 176 bp (mutated sample) were analyzed using Agilent 2100 Bioanalyzer. The 105 bp difference corresponds to the skipped exon 2. L: ladder; CTL-: negative control. ( C ) Screenshot of the Integrative Genomics Viewer (IGV) window presenting aligned RNA-seq data obtained from cases 744 (top) and a third T-ALL WT patient (693, bottom). The dotted line is centered on the donor splice site of exon 2 of MED12 (g.chrX:70339329). Case 744 shows aberrant splicing of exon 2, while 693 was WT for this position and shows a mature transcript that includes the exon 2.

    Article Snippet: Amplified fragments were analyzed on the Agilent 2100 Bioanalyzer Instrument and by Sanger sequencing (McGill University and Genome Quebec Innovation Centre).

    Techniques: Mutagenesis, Electrophoresis, Polymerase Chain Reaction, Synthesized, Isolation, Amplification, CTL Assay, Negative Control, RNA Sequencing Assay

    Comparison of small RNA extraction from MDA-MB-231 cells with fibers and columns analyzed by an Agilent 2100 Bioanalyzer. Small RNA recovery was as high as 985 pg/μL with the fibers and 10.2 pg/μL with the columns from as little as 134,000

    Journal: Analytical and bioanalytical chemistry

    Article Title: Extraction of microRNAs from biological matrices with titanium dioxide nanofibers

    doi: 10.1007/s00216-017-0649-3

    Figure Lengend Snippet: Comparison of small RNA extraction from MDA-MB-231 cells with fibers and columns analyzed by an Agilent 2100 Bioanalyzer. Small RNA recovery was as high as 985 pg/μL with the fibers and 10.2 pg/μL with the columns from as little as 134,000

    Article Snippet: The chip was run on an Agilent 2100 Bioanalyzer Instrument using the Small RNA Analysis Kit with 1 μL of extraction sample following the protocol provided by Agilent.

    Techniques: RNA Extraction, Multiple Displacement Amplification

    ( a ) Multiplex ligation-dependent genome amplification (MLGA), reaction scheme.  (I)  Genomic DNA is digested by restriction enzyme to generate targets with defined ends.  (II)  Each MLGA probe consists of two oligonucleotides, one selector oligo of 70–74 nt (green) and one general vector oligo of 34 nt (red). MLGA probe together with DNA-ligase forms circular DNA of target molecules after denaturation and hybridization.  (III)  To reduce background signal in the assay, undesirable, linear DNA is degraded by exonuclease I (Exo I).  (IV)  Multiplex PCR is facilitated by using universal primers that hybridize to a sequence in the vector. PCR products are analyzed using the Agilent Bioanalyzer 2100™ electrophoresis system. ( b ) Data from an MLGA set of 14 probes targeting loci on human chromosomes 13, 18, 21, X and Y. The upper graph shows the resulting elution diagrams from analyses of male and female DNA pools.

    Journal: Nucleic Acids Research

    Article Title: MLGA--a rapid and cost-efficient assay for gene copy-number analysis

    doi: 10.1093/nar/gkm651

    Figure Lengend Snippet: ( a ) Multiplex ligation-dependent genome amplification (MLGA), reaction scheme. (I) Genomic DNA is digested by restriction enzyme to generate targets with defined ends. (II) Each MLGA probe consists of two oligonucleotides, one selector oligo of 70–74 nt (green) and one general vector oligo of 34 nt (red). MLGA probe together with DNA-ligase forms circular DNA of target molecules after denaturation and hybridization. (III) To reduce background signal in the assay, undesirable, linear DNA is degraded by exonuclease I (Exo I). (IV) Multiplex PCR is facilitated by using universal primers that hybridize to a sequence in the vector. PCR products are analyzed using the Agilent Bioanalyzer 2100™ electrophoresis system. ( b ) Data from an MLGA set of 14 probes targeting loci on human chromosomes 13, 18, 21, X and Y. The upper graph shows the resulting elution diagrams from analyses of male and female DNA pools.

    Article Snippet: Temperature cycling was performed as follows: 37° C for 30 min, 95° C for 5 min followed by 30 cycles of 95° C 15 s, 55° C 30 s and 72° C for 60 s followed by 72° C for 10 min. PCR products were analyzed using an Agilent Bioanalyzer 2100™ instrument and quantified using the Agilent 2100 expert software, version B.02.02.SI238.

    Techniques: Multiplex Assay, Ligation, Amplification, Plasmid Preparation, Hybridization, Polymerase Chain Reaction, Sequencing, Electrophoresis