scholarly journals Restriction Digest v1 (protocols.io.bnwvmfe6)

protocols.io ◽  
2020 ◽  
Author(s):  
Zhujun Wei
Keyword(s):  
Restriction Digest

IDENTIFYING THE C677T POLYMORPHISM OF MTHFR GENE BY PCR-RFLP TECHNIQUE

2011 ◽  
pp. 28-35
Author(s):  
Keyword(s):  
Internal Control ◽  
Restriction Site ◽  
Mthfr Gene ◽  
C677t Polymorphism ◽  
Mthfr Genotype ◽  
Restriction Digest ◽  
Genotype Frequencies ◽  
Pcr Rflp ◽  
Sequencing Technique ◽  
Volunteer Group

Background: The C677T polymorphism of MTHFR gene is a risk factor of many diseases. This study is aimed at: (1) Improving a PCR-RFLP process with the own designed primers to identify the C677T polymorphism of MTHFR gene. (2) Evaluating the prevalence of the C677T polymorphism of MTHFR gene in volunteer group. Materials and method: DNA samples was extracted from peripheral blood of 60 volunteers. Designing primers by using FastPCR software, then improving PCR technique. Standardizing the optimal conditions of restriction digest by HinfI. Confirming the results of polymorphism by DNA sequencing technique. Results: We designed successfully primers to amplify fragment of MTHFR gene including C677T polymorphism and an obligatory restriction site of HinfI (as internal control). 0.5 µl of HinfI enzyme (10 U/µl) is enough for restriction digest. The MTHFR genotype frequencies were: 71.67 % (677CC); 25% (677CT); and 3.33 % (677TT). Conclusion: We standardized successfully PCR-RFLP technique to identifying C677T polymorphism of MTHFR gene. Keywords: C677T polymorphism, MTHFR gene, PCR-RFLP

Isotyping the Human TOMM40 Variable-Length Polymorphism by Gene Amplification and Restriction Digest

2012 ◽  
Vol 9 (10) ◽  
pp. 1168-1173
Author(s):  
Steven J. Greco ◽  
Ashkan Hamzelou ◽  
Jane M. Johnston ◽  
Sandy Richardson ◽  
Mark A. Smith ◽  
...  
Keyword(s):  
Gene Amplification ◽  
Length Polymorphism ◽  
Variable Length ◽  
Restriction Digest

Large‐Scale BAC Clone Restriction Digest Fingerprinting

2007 ◽  
Vol 53 (1) ◽  
Author(s):  
Carrie A. Mathewson ◽  
Jacqueline E. Schein ◽  
Marco A. Marra
Keyword(s):  
Large Scale ◽  
Bac Clone ◽  
Restriction Digest

Restriction Digest PCR (RD-PCR) for the Analysis of Gene Mutations. Application to Ki-ras

1998 ◽  
Vol 36 (8) ◽  
Author(s):  
Klaus Roland Huber ◽  
Jürgen Bittner ◽  
Kurt Bauer ◽  
Lorenz Trümper ◽  
Alexandra Sek ◽  
...  
Keyword(s):  
Gene Mutations ◽  
Restriction Digest

scholarly journals Long-Range Mapping of the Streptococcus agalactiae Phylogenetic Lineage Restriction Digest Pattern Type III-3 Reveals Clustering of Virulence Genes

2002 ◽  
Vol 70 (1) ◽  
pp. 134-139 ◽  
Author(s):  
John F. Bohnsack ◽  
April A. Whiting ◽  
Russell D. Bradford ◽  
Brenna K. Van Frank ◽  
Shinji Takahashi ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Streptococcus Agalactiae ◽  
Virulence Genes ◽  
Physical Map ◽  
Group B Streptococcus ◽  
Subtractive Hybridization ◽  
Chromosomal Dna ◽  
Type Iii ◽  
Phylogenetic Lineage ◽  
Restriction Digest

ABSTRACT Human isolates of serotype III Streptococcus agalactiae (group B streptococcus [GBS]) can be divided into three separate phylogenetic lineages based on analysis of the restriction digest patterns (RDPs) of chromosomal DNA. Nine DNA sequences that are present in all isolates of the RDP III-3 phylogenetic lineage, but not in the other lineages, were identified by genomic subtractive hybridization. A complete physical map of a III-3 chromosome was constructed. Six of the nine III-3-specific sequences mapped to a 340-kb Sse8387I fragment which contains or is located close to known GBS virulence genes. One of the III-3-specific probes, AW-10, encodes part of GBSi1, a group II intron that is inserted at two sites within the GBS genome. The second chromosomal site for GBSi1 was isolated, sequenced, and mapped to a location near the locus responsible for hemolysin production. These findings suggest that the genetic variation that distinguishes the RDP type III-3 strains from other serotype III strains occurs largely within localized areas of the genome containing known or putative virulence genes.

scholarly journals Restriction digest screening facilitates efficient detection of site-directed mutations introduced by CRISPR in C. albicans UME6

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e4920 ◽  
Author(s):  
Ben A. Evans ◽  
Olivia L. Smith ◽  
Ethan S. Pickerill ◽  
Mary K. York ◽  
Kristen J.P. Buenconsejo ◽  
...  
Keyword(s):  
Protein Function ◽  
Point Mutations ◽  
Pcr Amplification ◽  
Restriction Enzymes ◽  
Model Organisms ◽  
Efficient Manner ◽  
Restriction Digest ◽  
Efficient Detection ◽  
Wide Range ◽  
Restriction Sites

Introduction of point mutations to a gene of interest is a powerful tool when determining protein function. CRISPR-mediated genome editing allows for more efficient transfer of a desired mutation into a wide range of model organisms. Traditionally, PCR amplification and DNA sequencing is used to determine if isolates contain the intended mutation. However, mutation efficiency is highly variable, potentially making sequencing costly and time consuming. To more efficiently screen for correct transformants, we have identified restriction enzymes sites that encode for two identical amino acids or one or two stop codons. We used CRISPR to introduce these restriction sites directly upstream of the Candida albicans UME6 Zn2+-binding domain, a known regulator of C. albicans filamentation. While repair templates coding for different restriction sites were not equally successful at introducing mutations, restriction digest screening enabled us to rapidly identify isolates with the intended mutation in a cost-efficient manner. In addition, mutated isolates have clear defects in filamentation and virulence compared to wild type C. albicans. Our data suggest restriction digestion screening efficiently identifies point mutations introduced by CRISPR and streamlines the process of identifying residues important for a phenotype of interest.

scholarly journals The interaction between dam methylation sites and Xba1 restriction digest sites in Escherichia coli O157:H7 EDL933

2007 ◽  
Vol 102 (3) ◽  
pp. 820-825 ◽  
Author(s):  
J. Sales ◽  
L. Vali ◽  
D.V. Hoyle ◽  
C.M. Yates ◽  
S.G.B. Amyes ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Escherichia Coli O157 ◽  
Restriction Digest ◽  
Dam Methylation
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