Haplotype-Contained PCR Products Analysis by Sequencing with Selective Restriction of Primer Extension
نویسندگان
چکیده
منابع مشابه
Haplotype-Contained PCR Products Analysis by Sequencing with Selective Restriction of Primer Extension
We develop a strategy for haplotype analysis of PCR products that contained two adjacent heterozygous loci using sequencing with specific primers, allele-specific primers, and ddNTP-blocked primers. To validate its feasibility, two sets of PCR products, including two adjacent heterozygous SNPs, UGT1A1⁎6 (rs4148323) and UGT1A1⁎28 (rs8175347), and two adjacent heterozygous SNPs, K1637K (rs1117601...
متن کاملMultiplexed RNA structure characterization with selective 2'-hydroxyl acylation analyzed by primer extension sequencing (SHAPE-Seq).
New regulatory roles continue to emerge for both natural and engineered noncoding RNAs, many of which have specific secondary and tertiary structures essential to their function. Thus there is a growing need to develop technologies that enable rapid characterization of structural features within complex RNA populations. We have developed a high-throughput technique, SHAPE-Seq, that can simultan...
متن کاملAnalysis of primer-derived, nonspecific amplification products in RAPD-PCR.
Recently, we applied the randomamplified polymorphic DNA polymerase chain reaction (RAPD-PCR) (17), also known as arbitrarily primed PCR (AP-PCR) (16), procedure to fingerprint genomes of various varieties and wild relatives of sugarcane (Saccharum spp.) (4). Unlike restriction fragment length polymorphism (RFLP) (2), RAPD-PCR is a simpler, faster, less laborious and less expensive procedure. H...
متن کاملRapid sequencing of unpurified PCR products by thermal asymmetric PCR cycle sequencing using unlabeled sequencing primers.
Numerous methods have been developed for direct sequencing of single or double-stranded DNA amplified by PCR, and most require template purification steps prior to sequencing to remove excess unincorporated primers and dNTPs. Purification methods can be cumbersome, laborious and time consuming, or yield low DNA recoveries. Strategies for direct sequencing of doublestranded DNA products without ...
متن کاملAnalysis of RNA by primer extension.
1. Phosphorylate oligonucleotide probe: a. Oligonucleotide primer (5 – 7 pmoles or 60 ng) 1 μL b. dH2O 6.5 μL c. 10X kinase buffer 1.5 μL d. T4 PNK (~10 U) 1 μL e. [γ-P]ATP (7000 Ci/mmole) 2 μL 2. Stop the kinase reaction by adding 500 μL of TE (pH 7.6) 3. Add 25 μg of carrier RNA. 4. Phenol/Chloroform extract 2X 5. Ethanol precipitate with sodium acetate (pH 5.2) 6. Wash with 70% ethanol and r...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: BioMed Research International
سال: 2017
ISSN: 2314-6133,2314-6141
DOI: 10.1155/2017/1397902