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New England Biolabs
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About 2 products / 739 snippets / 42 images

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1 kb DNA Ladder
New England Biolabs  
Catalog#: N3232
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"no. 15593‐049) 3 M sodium acetate, pH 5.2 100% ethanol (Fisher, cat. no. 04355222) 10 mg/ml RNase A, DNase and protease free (Thermo Scientific, cat. no. EN0531) Agarose TBE (see recipe ) 1‐kb ladder (New England Biolabs, cat. no. N3232L) 6× Orange G gel loading dye (see recipe ) Dounce homogenizer (DWK Life Sciences, cat. no. 8853010002/8853030002) 1.7‐ml microcentrifuge tubes (Axygen, MCT‐175‐C)"  (see more)
98 / 100
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1 kb DNA Ladder
New England Biolabs  
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Catalog#: N3232
Techniques
 
Article Snippets
"no. 15593‐049) 3 M sodium acetate, pH 5.2 100% ethanol (Fisher, cat. no. 04355222) 10 mg/ml RNase A, DNase and protease free (Thermo Scientific, cat. no. EN0531) Agarose TBE (see recipe ) 1‐kb ladder (New England Biolabs, cat. no. N3232L) 6× Orange G gel loading dye (see recipe ) Dounce homogenizer (DWK Life Sciences, cat. no. 8853010002/8853030002) 1.7‐ml microcentrifuge tubes (Axygen, MCT‐175‐C)"  (see more)
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N IC A TI O N 3232
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"C O M M U N IC A TI O N 3232 www.advmat.de © 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim Adv."  (see more)
38+  more images
Elevation of viral genomic dsRNA accumulation levels by chromosomally expressed CHV1-EP713-encoded protein p29. (A) Agarose gel electrophoretic analysis of total RNA from mycelia of p29 transformants infected with p29 mutant viruses Δp29 and Δp69. As described in Materials and Methods, equal amounts (0.25 OD) of total RNA isolated from the mycelia of nontransformant (EP155), p29 transformant (Twtp29), or EGFP transformant (TEGFP) (43) that were infected with wild-type (wt), Δp29, or Δp69b (Δp69) mutant CHV1-EP713 were applied to each well of a 0.7% agarose gel and electrophoresed in a 1× TAE buffer system. M, <t>1-kb</t> <t>DNA</t> <t>ladder</t> standards (Gibco-BRL, Gaithersburg, Md). (B) Relative accumulation of viral minus strand-RNA in wild-type CHV1-EP713- and mutant virus-infected fungal colonies. Total RNA (shown in panel A) was isolated from untransformed (EP155) or p29-transformed (Twtp29) mycelia infected with wild-type CHV1-EP713 (wt) and mutant viruses Δp29 and Δp69b (Δp69) and used for strand-specific cDNA synthesis after denaturation in 90% dimethyl sulfoxide at 65°C (4). The resulting cDNA was subject to semiquantitative PCR analysis in a SmartCycler system (Takara) with cDNA of 18S rRNA generated in the same reverse transcriptase reaction for normalization. The sequences of primers and TaqMan probes used in the quantification were as described previously (44). Viral negative-strand RNA accumulation levels are reported as a percentage of the value measured for CHV1-EP713-infected colonies, with standard deviations indicated by the error bars based on three independent measurements.
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Deletion of adhE and PCR confirmation in T. saccharolyticum and C. thermocellum. Deletion of T. saccharolyticum adhE with P1 (external) and P2 (internal) confirmation primer pairs. (A) A resistance cassette is transformed into parent strain JW/SL-YS485, deleting adhE and creating strain LL1076. (B) T. saccharolyticum strain JW/SL-YS485 in lane 1 shows the expected 3.2-kb fragment with P1 external primers, and lane 3 shows the 900-bp internal P2 fragment of adhE. LL1076 in lane 2 shows a larger 5.3-kb band due to the replacement of 1.9 kb with the 4-kb disruption construct, and lane 4 shows the loss of the internal P2 band of adhE. (C) Deletion of C. thermocellum adhE with P3 external and P4 internal confirmation primer pairs. M1354 was transformed with plasmid pJLO19 and used to delete adhE to make strain LL1111. C. thermocellum strain M1354 in lane 1 shows the expected P3 external product size of 4.9 kb, and lane 3 shows the 800-bp P4 internal fragment. (D) LL1111 in lane 2 shows a 2.3-kb loss in adhE with a P3 product size of 2.6 kb, and lane 4 shows no P4 fragment. The DNA marker is the <t>1-kb</t> ladder from New England BioLabs and is marked with an L in panels B and D. Primer sequences can be found in Table 1.
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Agarose (1%) gel stained with ethidium bromide showing the PCR product amplified with the eap and map primers P2 and P3 from genomic <t>DNA</t> of S. aureus as described in Methods and Materials. Lanes: Mr, <t>DNA</t> <t>molecular</t> size marker (combined 100-bp and 1-kb DNA ladder [New England Biolabs]); 1, S. epidermidis DSM 20044; 2, S. aureus clinical isolate 7 (1.8 kb); 3, S. aureus Newman (2.0 kb); 4, S. aureus Wood 46 corresponding to group III (2.4 kb).
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Agarose gel electrophoresis profile of PCR product generated by using different sets of primers during the construction of hemolysin null mutant. Lanes 1 and 7 show the <t>1-kb</t> <t>DNA</t> <t>ladder</t> (New England Biolabs). Lanes 2, 3, and 4 show the PCR product generated from wild-type CO848b, plasmid pRM, and CO848b hlyA::tet, respectively, with primers hlyA (forward) and hlyA (reverse). Lanes 5 and 6 show the PCR product generated from CO848b (wild type) and CO848 (hlyA::tet), respectively, with primers hlyA (forward) and hlyB (reverse).
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