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Human Mitochondrial DNA - Amplification and Sequencing Standard Reference Materials - SRM 2392 and SRM 2392-1

Published

Author(s)

Barbara C. Levin, Diane K. Hancock, K A. Holland, He Cheng, Kristy L. Richie

Abstract

Standard Reference Materials SRM 2392 and 2392-I are intended to provide quality control when performing the polymerase chain reaction (PCR) and sequencing of human mitochondrial DNA (mtDNA) for forensic identification, medical diagnosis, or mutation detection. They may also serve as controls when amplifying (PCR) and sequencing any DNA. These SRMs can also be used for quality assurance when determining the sequence of in-house DNA controls. SRM 2392 is certified for the sequences of the entire human mtDNA (16,569 base pairs) from two lymphoblastoid cell culture lines (CHR and 9947A) from apparently normal individuals and the cloned HV1 region of CHR containing a C-stretch through which it is difficult to sequence. SRM 2392-I is certified for the mtDNA sequence from HL-60, a promyelocytic cell line prepared from the peripheral blood leukocytes from an individual with acute promyelocytic leukemia. The mtDNA sequence information (but not the DNA) of two additional DNA templates (GM03798 and GM10742A) that were amplified and sequenced in their entirety multiple times at NIST are provided in this Special Publication for comparison purposes. The sequences of fifty-eight unique primer sets that allow any area or the entire mtDNA (16,569 base pairs) to be amplified and sequenced are also provided.We found during the sequencing of these five mtDNA templates that some of the single nucleotide polymorphisms (SNPs) did result in amino acid changes when compared with the Cambridge Reference Sequence (Anderson et al.,1981; Andrews et al.,1999). Two interlaboratory evaluations, one for the amplification, sequencing, and data analysis of the CHR template and one for the HL-60 template were each conducted by three different laboratories and NIST. Corroboration of the results in SRMs 2392 and 2392-I will provide quality assurance that any unknown mtDNA is also being amplified and sequenced correctly.
Citation
Special Publication (NIST SP) - 260-155
Report Number
260-155

Keywords

9947A, CHR, forensic identification, GM10742A, GMO3798, HL-60, medical diagnosis, mitochondrial DNA sequence, mutation detection

Citation

Levin, B. , Hancock, D. , Holland, K. , , H. and Richie, K. (2003), Human Mitochondrial DNA - Amplification and Sequencing Standard Reference Materials - SRM 2392 and SRM 2392-1, Special Publication (NIST SP), National Institute of Standards and Technology, Gaithersburg, MD (Accessed May 18, 2024)

Issues

If you have any questions about this publication or are having problems accessing it, please contact reflib@nist.gov.

Created September 1, 2003, Updated October 16, 2008