Journal: Analytical chemistry

Article Title: Electrode-Free Concentration and Recovery of DNA at Physiologically Relevant Ionic Concentrations

doi: 10.1021/acs.analchem.0c00831

Figure Lengend Snippet: Microscopic snapshots of the DNA concentration process (a) with and (b) without HiSMRT. (a) Hind III digested λ DNA sample in 1× EB-HCl buffer was pumped into a reservoir of deionized (DI) water. At the outlet of the microcapillary, DNA molecules concentrated into a bolus with a growing size within 600 s of observation. (b) DI water was replaced by the 1× EB-HCl buffer to eliminate the ionic concentration. The absence of the ion concentration gradient resulted in dispersed DNA in the downstream of the DNA sample outlet without concentration (as shown in the dotted circle). This comparison confirms that an ionic concentration gradient is necessary for the HiSMRT DNA concentration.

Article Snippet: Hind III digested λ DNA (New England Biolabs) was used as the DNA sample for all experiments.

Techniques: Concentration Assay

Journal: Analytical chemistry

Article Title: Electrode-Free Concentration and Recovery of DNA at Physiologically Relevant Ionic Concentrations

doi: 10.1021/acs.analchem.0c00831

Figure Lengend Snippet: HiSMRT concentration experiment coupled with single-molecule-free solution hydrodynamic separation (SML-FSHS) on the cylindrical illumination confocal spectroscopy (CICS) platform for quantitative analysis of Hind III digested λ DNA. (a) Time trace of the single-molecule fluorescence bursts of DNA concentrated by HiSMRT, separated by SML-FSHS, and detected by CICS. (b) Free solution hydrodynamic separation chromatograms obtained by the single-molecule counting analysis provides quantitative capabilities for the concentration performance. It shows that many DNA molecules are detected with HiSMRT, while only a low level of random noise was detected without HiSMRT. These results confirm that HiSMRT indeed concentrated DNA and verify the observation on the microscope in Figure 2.

Article Snippet: Hind III digested λ DNA (New England Biolabs) was used as the DNA sample for all experiments.

Techniques: Concentration Assay, Spectroscopy, Fluorescence, Single Molecule Counting, Microscopy

Journal:

Article Title: Molecular Analysis of Mycobacterium avium Isolates by Using Pulsed-Field Gel Electrophoresis and PCR

doi:

Figure Lengend Snippet: Restriction patterns from AseI digests of M. avium isolates resolved by PFGE. Lanes: 1 and 6, bacteriophage lambda DNA concatemers (sizes [in kilobases] are indicated on the left); 2, isolate 100A8; 3 and 4, pattern P7 (isolates 100A28 and 100A32, respectively); 5, isolate 100A25; 7 to 11, five sequential isolates from one patient, respectively (pattern P1); 12 to 15, four isolates from two patients, respectively (pattern P2).

Article Snippet: Lanes 6 and 18, bacteriophage lambda DNA- Bst EII digest; lane 12, pBR322 DNA- Msp I digest (New England Biolabs); lanes 1 to 5, five sequential isolates from one patient, respectively (PCR profile A, PFGE pattern P1); lanes 7, 8, and 9, three sequential isolates from one patient, respectively (PCR profile A, PFGE pattern P2) (no amplification product was detected in lane 8 in that experiment); lane 10, isolate 100A31 from another patient (PCR profile A, PFGE pattern P2); lanes 11 and 13, two sequential isolates from one patient, respectively; lane 14, isolate 100A7 from a different patient (PCR profile E, PFGE pattern P6 or a unique pattern); lanes 15, 16, and 17, isolates 100A13, 100A26, and 100A30, respectively.

Techniques: Lambda DNA Preparation

Journal:

Article Title: Molecular Analysis of Mycobacterium avium Isolates by Using Pulsed-Field Gel Electrophoresis and PCR

doi:

Figure Lengend Snippet: PCR typing of clinical M. avium isolates. Lanes: 1 to 5, 7 to 11, and 13 to 17, patterns of isolates obtained from 15 unrelated patients; lanes 6 and 18, bacteriophage lambda DNA-BstEII digest molecular weight marker; and lane 12, pBR322 DNA-MspI digest (New England Biolabs).

Article Snippet: Lanes 6 and 18, bacteriophage lambda DNA- Bst EII digest; lane 12, pBR322 DNA- Msp I digest (New England Biolabs); lanes 1 to 5, five sequential isolates from one patient, respectively (PCR profile A, PFGE pattern P1); lanes 7, 8, and 9, three sequential isolates from one patient, respectively (PCR profile A, PFGE pattern P2) (no amplification product was detected in lane 8 in that experiment); lane 10, isolate 100A31 from another patient (PCR profile A, PFGE pattern P2); lanes 11 and 13, two sequential isolates from one patient, respectively; lane 14, isolate 100A7 from a different patient (PCR profile E, PFGE pattern P6 or a unique pattern); lanes 15, 16, and 17, isolates 100A13, 100A26, and 100A30, respectively.

Techniques: Lambda DNA Preparation, Molecular Weight, Marker

Journal:

Article Title: Molecular Analysis of Mycobacterium avium Isolates by Using Pulsed-Field Gel Electrophoresis and PCR

doi:

Figure Lengend Snippet: Electrophoretic PCR patterns for M. avium isolates. Lanes 6 and 18, bacteriophage lambda DNA-BstEII digest; lane 12, pBR322 DNA-MspI digest (New England Biolabs); lanes 1 to 5, five sequential isolates from one patient, respectively (PCR profile A, PFGE pattern P1); lanes 7, 8, and 9, three sequential isolates from one patient, respectively (PCR profile A, PFGE pattern P2) (no amplification product was detected in lane 8 in that experiment); lane 10, isolate 100A31 from another patient (PCR profile A, PFGE pattern P2); lanes 11 and 13, two sequential isolates from one patient, respectively; lane 14, isolate 100A7 from a different patient (PCR profile E, PFGE pattern P6 or a unique pattern); lanes 15, 16, and 17, isolates 100A13, 100A26, and 100A30, respectively.

Article Snippet: Lanes 6 and 18, bacteriophage lambda DNA- Bst EII digest; lane 12, pBR322 DNA- Msp I digest (New England Biolabs); lanes 1 to 5, five sequential isolates from one patient, respectively (PCR profile A, PFGE pattern P1); lanes 7, 8, and 9, three sequential isolates from one patient, respectively (PCR profile A, PFGE pattern P2) (no amplification product was detected in lane 8 in that experiment); lane 10, isolate 100A31 from another patient (PCR profile A, PFGE pattern P2); lanes 11 and 13, two sequential isolates from one patient, respectively; lane 14, isolate 100A7 from a different patient (PCR profile E, PFGE pattern P6 or a unique pattern); lanes 15, 16, and 17, isolates 100A13, 100A26, and 100A30, respectively.

Techniques: Lambda DNA Preparation, Amplification