Reagent Kits

AATI's capillary electrophoresis instruments utilize specially designed reagent kits to analyze diverse nucleic acid samples, from small RNA to genomic DNA, and NGS library quality control.

Reagent Kit Box

FP-1002-0275

gDNA 165Kb Analysis Kit

AccuCleave Agarose Kits
gDNA 165Kb Analysis Kit

The gDNA 165Kb Analysis Kit enables the separation of gDNA and DNA fragments through 165Kb at low concentrations. Replace PFGE with pulsed-field capillary electrophoresis at critical QC checkpoints to conserve sample and obtain results faster.

Components

275 Sample Kit
Large DNA Separation 5001 Gel 250 mL (FP-5001-0250)
FEMTO Pulse Intercalating Dye 30 µL (FP-6001-U030)
5X Inlet Buffer 75 mL (DNF-325-0075)
5X Conditioning Solution 50 mL (DNF-425-0050)
gDNA Diluent Marker 3 mL x 2 vials (FP-8001-0003)
165Kb Ladder 200 pg/µL 75 µL (FP-7002-U075)
0.25X TE Rinse Buffer 60 mL (DNF-497-0060)
BF-P25 Blank Solution 5 mL (DNF-306-0005)
Storage Solution 100 mL (GP-435-0100)
Eppendorf LoBind® 0.5 mL tubes package of 50

Specifications

Specifications Description
Sample Volume Required 2 µL
Total Electrophoresis Run Time 70 min (22-40 Array)
DNA Sizing Range 1.3 Kb – 165 Kb
Sizing Precision 20% CV
Sizing Accuracy ± 15%
DNA Fragment Concentration Range 0.3 pg/µL – 30 pg/µL input DNA
DNA Smear Concentration Range 5 pg/µL – 500 pg/µL input DNA
Maximum gDNA Concentration 500 pg/µL
gDNA Quantification Precision 25% CV

Examples

Quantify and qualify gDNA smears and fragments through 165 Kb with the gDNA 165Kb Analysis Kit (FP-1002-0275) for the FEMTO Pulse™ Automated Pulsed-Field CE Instrument. A high quality DNA ladder is essential to the accurate sizing and quantification of large DNA smears and fragments. The 165Kb Ladder (FP-7002-U075) is composed of DNA fragments at concentrations suitable to create robust standard curves for sizing and quantification (Figure 1).

Figure 1
Figure 1. Separation of the 165Kb Ladder 200 pg/µL (FP-7002-U075) performed on the FEMTO Pulse Automated Pulsed-Field CE Instrument equipped with an Ultra-Short Capillary Array (22-40). The gDNA 165Kb Analysis Kit was used.

Unprecedented sizing and detection sensitivity of large gDNA smears and fragments are achieved with the gDNA 165Kb Analysis Kit (FP-1002-0275). As genomics and molecular biology have progressed, the ability to qualify and quantify large DNA samples at low concentrations has grown more important, most notably for long read sequencing. gDNA smears are readily sized through 165 Kb and quantified as low as 5 pg/µL.

Smear Analysis and the Genomic Quality Number (GQN) are two analytic functions provided in PROSize Data Analysis Software to simplify the post-electrophoretic analysis of gDNA smears. Designated by the user and defined by dotted orange lines on the electropherogram (Figure 2), Smear Analysis enables a more inclusive analysis of gDNA smears than standard Peak Analysis. Similarly, the GQN is a dynamic, user-defined quality metric. To employ the GQN, users first define a size threshold, graphically depicted as a dotted blue line on the electropherogram (Figure 2). PROSize then determines the GQN based upon how much of the sample exceeds the user-selected size threshold, assigning a numeric value between 0 and 10.

Figure 2
Figure 2. Separation of commercially prepared sheared gDNA using a FEMTO Pulse Automated Pulsed-Field CE Instrument equipped with an Ultra-Short Array (22-40). The upper panel shows an electropherogram of a gDNA sample diluted to 8 pg/µL. The bottom panel shows an electropherogram of gDNA diluted to 500 pg/µL. Dotted orange lines delineate the Smear Analysis region. The dotted blue line illustrates the GQN size threshold.

Large fragment DNA analysis is important to a plethora of applications in molecular biology, including the construction of large genetic elements. For such applications, sample conservation and an extended sizing capability are both essential. Large DNA fragments, like lambda DNA, are quantified down to 300 fg/µL and sized through 165 Kb (Figure 3).

Figure 3
Figure 3. Separation of commercially prepared lambda genomes using a FEMTO Pulse Automated Pulsed-Field CE Instrument equipped with an Ultra-Short Array (22-40). The upper panel depicts an electropherogram of a lambda genome diluted to 500 fg/µL. Similarly, the bottom panel depicts an electropherogram of the separation of a lambda genome diluted to 16 pg/µL.
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