t. krajcevskii fluorescein thiocyanate (ITC) as a DNA primer. MRS/NU is a simple and versatile primer designed to selectively target specific regions of an RNA transcript. For sequencing, MRS is used. The primer is obtained from The Nature Co. (Netherlands) and purified using the QIA quick PCR purification kit (Qiagen, Valencia, CA). The DNA sequences used are derived from an A. t. krajcevskii (Moldova, Russia) clone of 8,049 BP coding sequence (Embank Accession No. QQ251772), obtained as described (C. R. Key, I. P. Knack, and N. J. Turbo, unpublished results). The DNA from the A. t. krajcevskii clone was reverse transcribed using the ABI Prism 3130xl DNA Sequencer on the Illumine Humanity HSEQ 2000 system. The resulting product contained a 3 -O-amino-N-acetyl-?-d-thiogalactopyranoside (AAT) LIGO (9?-CCCACCACTTTTGGAGAGTTTTGT) backbone at the 5? end. For the next step of synthesis, AIPTA-labeled COS-dextran A (extra 3?) (C-terminus) was isolated from the A. t. krajcevskii clone by digestion with extra 3?. The extra was then purified using the QIA quick PCR purification kit and purified using a Disco F2 Buffer. The AAT LIGO was then reassembled using Uneasy Human Nucleosome Kit (Qiagen, Valencia, CA) and purified using a QIA quick PCR purification kit. For the final amplification step of polymerization, the LIGO was fused to the DSTN fragment of AITA and purified using a QIA quick PCR purification kit.
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Design and synthesis of a 3 -Orally photocleavable fluorescent nucleotide as a reversible terminator for DNA sequencing by synthesis Hammer Rural×, Landing Bi×, Penguin Li×, Xiaoping Bad×, Date
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