November 4, 2015

Molecular Biology Research

Specifying your project's requirements correctly, we provide you the following services of molecular biology techniques.
Please click on the titles for more information

Providing free consultancy for project proposal application, if you get one our research service in your project.

SNP GENOTYPING SERVICE

1. Sanger Sequencing Analysis:

This is as well-known gold standard method for identification of any mutation or polymorphism in target genomic sequence. Process includes the fluorescent labelled ddNTP incorporation during single strand synthesis and the products are read by using capillary electrophoresis device. The most efficient and maximum length is accepted to be 600 bases read.

You may prefer Sanger sequencing if your application is among the below subjects:

- Sequence analysis of unknown target genomic regions, De-novo sequencing, varification of NGS data, mitochondrial dna sequencing etc,

- HLA typing,
- Identification of unknown genomic variations related to a disease or protein aberrations
- Forensics
- Agriculture and Animal Breeding
- Identification of epigenetics related regions on DNA and methylation analysis
- Identification of microorganisms related pathogenic diseases or environmental conditions by 16S rRNA gene sequence analysis and metagenomics

Please get in contact if you are interested about this method.

 2. Restriction Endonuclease Analysis (REA):</b>

Type-2 Restrion Endonuclease enzymes bind on and cut double-stranded DNA sequences with a sequence specific manner. They first discovered as an enzyme of bacterial defence mechanism against phages and then widely accepted in genome engineering. Restriction enzymes are both used in gene cloning and SNP genotyping applications.

Sanger sequencing is required for finding unknown genomic variations at specific locations, however it is expensive and time-consuming for scanning of large number of samples for a specific point. In this case, after amplification of target sequence by PCR, variations can be identified by observation of PCR products as cut or un-cut by specific restriction enzymes.

Experimental set-up is crucial for obtaining correct results. For getting best we suggest the preliminary preparation as below:

Point mutation can be found by either cutting polymorphic allele or normal allele via enzymes. Since type-2 enzymes function with a sequence specific manner, one should check both possibilities to use best and cheaper enzyme.

For finding the enzymes according to target seqeunces, the free-online tool option is NEB Cutter (http://nc2.neb.com/NEBcutter2/)

The enzyme to be used should only cut the PCR product at the presence or at the absence of variation. Otherwise, the variation can not be discriminated.

Target PCR products may possess more than one similar sequences, so in order to eliminate multiple restriction of sequences, one should check the presence of multipile recognition sites of enzymes on target sequence. If there is not, perfect.

The mutation/polymorphism point should not be at the central region of PCR product sequence. After cutting by enzyme, the result is observed as two seperate lines. Wrong data may be got by observing single line at every sample unless one optimize primers of PCR.

Please get in contact if you are interested about this method.

3. Allele Specific PCR Analysis:

Scanning of specific variations on samples can not be performed by Restriction Enzyme Assay because there is not enough identified type-2 endonucleases for all sequence possibilities. Additionally, same enzyme binding sequences may be present at very close to target variation. In this case REA wont work.

Fast SNP genotyping can also be performed by an alternative method called Allele-Specific PCR. This method uses specific primers of which 3′-end hybridizes on the polymorphic/mutant nucleotide. By making two PCR reactions with 2 forward primers and one cummon reverse primer, one can find the mutation by observing the PCR product lines on gel.

Experimental set-up is crucial for obtaining correct results. For getting best we suggest the preliminary preparation as below:

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Please get in contact if you are interested about this method.

PROTEIN ANALYSIS SERVICE

1. Surface Plasmon Resonance (SPR) Analysis

The theory is based on a physical process of light-metal film interaction. In SPR device, a material that has a high refractive index (with respect to medium) is coated with thin layer gold. When this gold coated material is treated with monochromatic light by a certain angle, some of the energy of light is transferred to free electrons of the gold and surface plasmons are formed on the gold. This plasmon will end up with specific refractive index. This phenomenon has been used for any molecular detection.

The application of SPR: On thin film gold surface, the molecule of interest is immobilized. This can be antibody, aptamer, cell, enzyme or etc. When the target molecule is sent to the media on top of molecule of interest and when association or dissociation happens between the immobilized olecule and target analyte, the refractive index will be changed which we can measured with high precision in real time by embedded sensors.

SPR technique has many advantages and label free detection is the most important advantage of it. SPR can be used to analyze biological or chemical materials and their molecular interactions without labeling molecules.

Real-Time analysis can be achieved with SPR techniques. Avidity and affinity of the samples can be detected. SPR technique offers almost at same level sensivity with ELISA and sometimes better.

Quantitave Analysis is an important feature with SPR devices. Every biological and chemical interaction can be monitored in details.

Please get in contact if you are interested about this method.

2. SDS-PAGE, 2D Gel Electrophoresis, and Electroblotting

This service is not active yet. Our plan is to make it available on next summer. Working hard on this.

Please get in contact if you are interested about this method.

3. ELISA Analysis

This service is not active yet. Our plan is to make it available on next summer. Working hard on this.

Please get in contact if you are interested about this method.

 

EPIGENOMICS ANALYSIS SERVICE

1. CpG Methylation and Hydroxymethylation Scanning

Please get in contact if you are interested about this method. More information will be available soon here.

2. Methylation Specific Restriction Endonuclease Analysis

Please get in contact if you are interested about this method. More information will be available soon here.

3. Methylation Specific PCR

Please get in contact if you are interested about this method. More information will be available soon here.

APTAMER SELECTION SERVICE

1. SELEX for Aptamer Identification

Please get in contact if you are interested about this method. More information will be available soon here.

 

GENE MODIFICATIONS SERVICE

1. CRISPR-Cas9 Application

This service is not active yet. Our plan is to make it available on next summer. Working hard on this.

Please get in contact if you are interested about this method.

2. Plasmid Designing, Cloning, and Bacteria Transformation

This service is not active yet. Our plan is to make it available on next summer. Working hard on this.

Please get in contact if you are interested about this method.

3. siRNA – miRNA Designing and Transfection

This service is not active yet. Our plan is to make it available on next summer. Working hard on this.

Please get in contact if you are interested about this method.

IMMUNOFLUORESCENCE / APTAMER-FLUORESCENCE SERVICE

1. Immunofluorescence Application

Please get in contact if you are interested about this method. More information will be available soon here.

2. Aptamer-Fluorescence Application

Please get in contact if you are interested about this method. More information will be available soon here.