ApaLI NEB#R0507

Restriction Enzymes ApaLI / Alw44I

Experiment
Restriction Enzymes ApaLI / Alw44I
Product
ApaLI NEB#R0507 from New England BioLabs
Manufacturer
New England BioLabs

Protocol tips

Protocol tips
PCR products were digested with NdeI, ApaLI, and AccI (NEB) at 37°C, ethanol-precipitated, and run on denaturing polyacrylamide gels as described in ref. 15.

Publication protocol

Superscript II (Invitrogen) reverse transcription reactions, performed according to the supplied protocol, contained 5–10 μg of RNA and reverse primer GAATTCTAACCCCTGTGCTAGCGCTT for tissue culture experiments or a nested primer, ATGCTTTCAAGCATCTGACCTAACC, for mouse experiments. In all cases, 20% of the reverse transcription reaction was PCR-amplified by using Taq (Qiagen, Valencia, CA) with sense primer GAATTCCCAGACGTCGCTGCATGC and antisense primer GAATTCTAACCCCTGTGCTAGCGCTT. The sense primer was end-labeled by using T4 polynucleotide kinase (NEB, Beverly, MA) and was used at a 1:5 ratio with the unlabeled sense primer (15). PCR products were digested with NdeI, ApaLI, and AccI (NEB) at 37°C, ethanol-precipitated, and run on denaturing polyacrylamide gels as described in ref. 15.

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Manufacturer protocol

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