Molecular Biology Services

1.Gene Cloning

Sangon Biotech have developed particular technology for obtain DNA fragments ("gene") with specific functions either by cloning from natural existing organism or by chemical gene synthesis. Cloning is usually more convenient than gene synthesis, but cloning is also time-consuming and sometimes can be difficult. Sangon Biotech not only help you to obtain the desired gene, but also help you to subcloning the desired gene into any vectors to save your time and money.

  • Direct gene cloning with known sequence from known source
  • Norvel gene cloning using RACE (Rapid Amplification of cDNA Ends) technology

2.Gene Expression Analysis Using Real-time Fluorescent Quantitative RT-PCR

Gene expression is the process by which information from a gene is used in the synthesis of a functional gene product (either proteins or non-protein coding genes such as miRNA). The regulation of gene expression process was strictly controlled in a complicated network in vivo. Gene expression profiling indicates the role of a particular gene and the changes in gene expression is associated with many biological process such as disease developing. So monitor the changes in gene expression becoming an more and more important method to monitor the "action" of particular genes.

Real-time PCR analysis is the gold standard for quantifying gene expression. We have developed methods for sensitive, accurate quantification of mRNA or microRNA using real-time PCR. Both TaqMan probe-based analysis and SYBR Green dye-based analysis are available. Individual samples or 96 or 384 well plates can be analyzed.

  • mRNA Real Time Fluorescent Quantitative RT-PCR
  • microRNA Real Time Fluorescent Quantitative RT-PCR

3.Genotyping

Microsatellite (STR) Genotyping

Microsatellites, also known as short tandem repeats (STR), are widely used molecular markers in genetics. Sangon Biotech offers a complete microsatellite (MS) genotyping service.

Single Nucleotide Polymorphism (SNP) Genotying

SNPs are one of the most common types of genetic variation which is associated with many diseases. SNP genotyping is the measurement of genetic variations of single nucleotide polymorphisms (SNPs) between members of a species. Sangon Biotech have developed several SNP genotyping platforms for different requirement including:

  • Direct Sequencing
  • Restricted Fragment Length Polymorphisms (RFLP)
  • TaqMan Probe
  • Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS)

4.cDNA Library Construction & EST Sequencing

cDNA library construction is a powful tool for gene expression, gene function research, norvel gene characterization, and many other applications. But obtain high-quality cDNA libraries is time-consuming and can be very challenging. Sangon Biotech has over 10 years of experience in generating cDNA libraries from various of samples. Thanks to our comprehensive sequencing platform. cDNA libraries with ESTs (expressed sequence tag) can also be sequenced and analysized here.

5.Species identification & Microbial Population Analyzing

  • Identification of Bacteria or Fungi
  • PCR-DGGE (Denaturing Gradient Gel Electrophoresis) analysis for the identification of microbial populations of Bacteria
  • Microbial Population Analyzing

6.Suppression Subtractive hybridization(SSH) Service

7.Single Nucleotide Polymorphism(SNP) genotying Service

SNP service content price and procedure

SNP testing service platform

  • Direct sequencing
  • Restricted fragment length polymorphisms (RLFP)
  • TaqMan Probe
  • MALDI-TOF MS

8.Illumina beadchip Services

  • Illumina beadchip SNP genotyping
  • Illumina beadchip methylation
  • Illumina beadchip gene expression profiles
  • Illumina beadchip microRNA expression profiles

9.Methylation Analysis

  • Bisulfite Sequencing Method (BSP)
  • Methylation Specific PCR Method (MSP)

10.Virus packaging

  • Adenovirus packaging services
  • Lentiviral packaging services

11.RNA Interference (siRNA) Experimental Services

1.Gene Cloning

Services Price ($) Turnaround Time Remarks
Known gene cloning from Prokaryotic or eukaryotic genome Minimum charge at $80 (<=1,000 bp)
For gene length >1,000 bp, $0.1 / extra bp
15 business days Extra charge for high GC
Known gene cloning from Eukaryotes cDNA Minimum charge at $130 (<=500 bp)
For gene length >500, $0.17 / extra bp
20 business days Extra charge for high GC
5'RACE Minimum charge at $800 (<=500 bp)
For fragments >500 bp, $0.34 / extra bp
1 to 2 months Including 3-5 designs of specific primers, RACE PCR and cloning sequencing.
3'RACE Minimum charge at $500 (<=800 bp)
For fragments >800 bp, $0.17 / extra bp
1 to 2 months

All data (including gel analysis photos of PCR, Sequencing trace files, etc) will be provided.

2.Gene Expression Analysis

Project Quantity Price ($)
Primer charge mRNA Pair $8.5/pair
miRNA Pair $13.5/pair
Probe Probe $165/probe
DNA extraction Blood sample $3.35/sample
Cells, animal tissues, etc. sample $5/sample
Water, mud, soil samples sample $5~$8.35/sample
RNA extraction + reverse transcriptase sample $13.5/sample
plasmid construction (required by Absolute quantification method) plasmid $16.5/plasmid
Standard Curve** 2 repeats #gene* 5 point*2 repeat $5/reaction
3 repeats #gene* 5 point*3 repeat $5/reaction
qPCR of Samples 1 repeats #Sample * #gene *1 repeat $5/reaction
2 repeats #Sample * #gene* 2 repeat $5/reaction
3 repeats #Sample * #gene* 3 repeat $5/reaction

**By default, 5 gradient dilution is used.

3.Microsatellite (STR) Genotyping

Steps Content Price ($)
DNA extraction Whole blood samples $3.5 /sample
Paraffin sections, tissue samples $8.5 /sample
Pre-test optimize PCR reaction conditions $33.5 for each amplified region
PCR reaction According to preliminary results of a large number of PCR Tube / $0.85 ~ $1.7
(based on the difficulty of PCR)
Label primers Synthesis of fluorescent labeled primers $50~$83.5/oligo
Formal test STR genotyping using ABI 3730XL DNA analyzer $250 /96 reaction
(without internal standard), minimum charge: 96 reaction
Requirements for samples Genomic DNA Concentration > 20 ng/μl, amount > 20 μl
(Specific amounts required according to experimental requirements)
Whole blood samples 1 ml anticoagulated blood
(please use EDTA as anticoagulant)
Paraffin sections, tissue samples >100 mg
Experimental period 4-6 weeks (Specific time accord to STR loci /sample number)

All data including excel table with site statistics situation, sequencing trace files and original documents will be provided.

4.Library Construction

Project Content Price ($)
SMART cDNA library cDNA library construction $2500 (provide 20 sequencing reactions for free)
Normalized cDNA library Normalized cDNA library construction $13,000 (provide 20 sequencing reactions for free)

EST sequencing

<5000 clones $3.2 per clone
5000-10000 clones $3.7 per clone
>10000 clones Inquire
Standard bioinformatics analysis Free
Advanced bioinformatics analysis Inquire
Sample Requirement High quality samples are required,
Tissue: >1 g
Cultured cells: >1 * 107
total RNA: > 20μg
Experimental period 4-6 weeks

5.Strain indenification & Microbial Population analyzing Services

5.1Strain identification

a.Services

  • Full-length 16S rDNA sequencing: The full-length 16S rDNA (~ 1,500 bp) was amplified and cloned into cloning vector by TA-cloning and followed by Sanger sequencing. More comprehensive information could be obtained by analyzing full-length 16S rDNA sequence.
  • Fungal ITS analysis: Identification of fungal by sequence of ITS (Internal transcribed spacer) region..
  • Fungal 18S rDNA analysis: Similar with 16S rDNA analysis of bacteria, 18S rDNA of Fungal is widely used for species identification.
  • Fungal 26S rDNA analysis: Particularly suitable for yeast strain identification.

b. Fees

Services Sample Timeline
Genomic DNA Bacteria
6s rDNA Sequencing $50 /sample
(Cloning and sequencing)
$65/sample
(Cloning and sequencing)
7 ~ 15 business days
$33.5/sample
(PCR products are sequenced)
$50/sample
(PCR products are sequenced)
Fungal ITS Sequencing $50 /sample
(Cloning and sequencing)
$85/sample
(Cloning and sequencing)
7 ~ 15 business days
$33.5/sample
(PCR products are sequenced)
$65/sample
(PCR products are sequenced)
Fungal 18s rDNA Sequencing $50 /sample
(Cloning and sequencing)
$100/sample
(Cloning and sequencing)
7 ~ 15 business days
$33.5/sample
(PCR products are sequenced)
$65/sample
(PCR products are sequenced)
Fungal 26S rDNA D1 / D2 region Sequencing $50 /sample
(Cloning and sequencing)
$65/sample
(Cloning and sequencing)
7 ~ 15 business days
$33.5/sample
(PCR products are sequenced)
$50/sample
(PCR products are sequenced)

c.Results

All related data (including photos of gel analysis, NCBI reference strain identification results, etc.), sequencing trace files will be provided.

5.2 Microbial Flora analysis

Services Sample Timeline Remarks
Genomic DNA Bacteria
RAPD $8.5/sample $25/sample 10 to 15 business days /
Rep-PCR $8.5/sample $25/sample 15 to 20 business days /
PCR-RFLP $13.5/sample $30/sample 15 to 20 business days /
PCR-SSCP $16.5/sample $33.5/sample 15 to 20 business days /
ARDRA $83.5/sample
(mixed bacteria)
$100/sample
(mixed bacteria)
20 to 25 business days Recovery and cloning of difference bands
Sequencing charge of $80/clone
LARD 833.5 /sample(mixed bacteria) 20 to 25 business days Including 50 positive clones
Partial sequence analysis
DGGE 250 /sample(mixed bacteria) 25 to 30 business days Recovery and cloning of difference bands
Sequencing charge of $80/clone

c. Results

  • The report, including material and methods, photo of gel analysis.
  • Construction of RIS Library, sequencing trace files, the plasmid containing the gene and puncture bacterial.
  • data analysis, including the NTSYS-pc software for fingerprint cluster analysis, phylogenetic tree constructed maps.

6.Suppression Subtractive hybridization

price and experimental period: 5835 dollar,45 working days

7.Single Nucleotide Polymorphism (SNP) Genotying Services

Typing methods Quantity Unit price Timeline Remarks
Mutation or SNP screening by Sanger sequencing <=20 exons $13.5/exon 10 business days Charge of primers will be calculated case by case
20~100 exons $10 /exon 14 business days >100, Inquire
Sequencing varification for known SNPs <=20 sites $10/site 10 business days >100, Inquire
20~100 sites $8.5/site 14 business days
PCR-RFLP <=500 sites $2 /site 15 business days >1,000, Inquire
500~1,000 sites $1.7/site 25 business days
TaqMan probe <=500 sites $3.5/site 20 business days Charge of primers will be calculated case by case
>1,000, Inquire
500~1,000 sites $3 /site 25 business days
Multiplex
SNaPshot
The following price is for number of samples <=500 For samples>500, SNP site>20 per sample, Inquire
<=2 sites per sample $8.5 sample 30 business days
3~10 sites per sample $1.7~$3.5/site 30 business days
11~20 sites per sample $0.85~$1.7/site 30 business days
MALDI-TOF The following price is for number of samples <=1000 For samples>1000, SNP site>20 per sample, Inquire
<=4 sites per sample $17 /sample 30 business days
5~10 sites per sample $1.5~2 /site 30 business days
11~20 sites per sample Inquire Inquire
LDR 3~10 sites per sample $1~2/site 15 business days samples>100
charge of primer will be calculated case by case
>11 sites per sample <=$1/site 20 business days

8.Illumina fiber beads confocal analysis platform for high throughput gene

8.1 Illumina Fiber beads chip price of services

Type Style Price($) Remarks
Whole-genome SNP genotyping chip HumanCytoSNP-12 v1.0 DNA Analysis BeadChip Kit 416.7 /case A multiple of 12 samples
Human660W-Quad v1.0 DNA Analysis BeadChip (16) 933.5/case A multiple of 16 samples
Human1M-Duo v2.0 DNA Analysis Kit (8) 1466.7 /case A multiple of 8 samples
CanineSNP20 Canine whole genome SNP genotyping chip 666.7 /case A multiple of 48 samples
BovineSNP50 Bovine whole genome SNP genotyping chip 633.5 n/case A multiple of 48 samples
OvineSNP50 Sheep genome SNP genotyping chip 583.5 /case A multiple of 48 samples
PorcineSNP60
Porcine whole-genome SNP genotyping chip
583.5 /case A multiple of 48 samples
Linkage analysis of SNP chip Linkage Panel Linkage analysis of human chip 250 /case A multiple of 96 samples
Mouse LD Linkage Linkage analysis of low-density chips mice 250 /case A multiple of 96 samples
Mouse MD Linkage Chip density linkage analysis in mice 416.7/case A multiple of 96 samples
  Whole genome expression microarray HumanRef-8 BeadChip Expression of the human genome chip 533.5 /case Best of 8 samples
Human-6 BeadChip Expression of the human genome chip 1083.5 /case Best of 6 samples
MouseRef-8 BeadChip Mouse whole genome expression chip 833.5 /case Best of 8 samples
Mouse-6 BeadChip Mouse whole genome expression chip 1083.5 /case Best of 6 samples
RatRef-8 BeadChip Rat whole genome expression chip 833.5 /case Best of 12 samples
MicroRNA chip Human microRNA expression profiling
microarray
1041.7 /case Best of 12 or 96 samples
mouse microRNA expression profiling microarray 1041.7 /case
Methylation microarray Human Methylation27
Methylation of the human genome chip
1041.7 /case At least 12 samples
Methylation Cancer Panel I Methylation in human cancer chip 833.5 /case

Type Style Sites Price(dollar /case) samples/ chip
Human genome SNP genotyping chip Human Omni5-Quad (new) 5000,000 2,000 4 samples
Human Omni2.5-8 2500,000 1333.5 8 samples
Human Omni1-Quad 1140,000 916.7 4 samples
Human OmniZhongHua-8 (new) 900,000 750 8 samples
Human OmniExpress-12 700,000 666.7 12 samples
Human CytoSNP-12 300,000 583.5 12 samples
Human Exome-12(new) 250,000 366.7 12 samples
Human Linkage-24 6056 533.5 24 samples
Animal plant genome SNP genotyping chip Cow HD chip 70 800 8 samples
Cow SNP50 chip 50,000 666.7 24 samples
Cow LD chain chip 6000 366.7 24 samples
Dog HD chip 170,000 800 12 samples
Sheep SNP50 chip 50,000 666.7 12 samples
Horse SNP50 chip 50,000 666.7 12 samples
Pig SNP60 chip 60,000 666.7 12 samples
Corn SNP50 chip 50,000 666.7 24 samples
Methylation detection chip Human Methylation 27K 27,578 833.5 12 samples
Human Methylation 450K 480,000 1133.5 12 samples
Microarray HumanHT-12 V4 47,231 633.5 12 samples
MouseWG-6 45,281 700 6 samples
Custom SNP chip(more than 480 samples)
Sites 96 144 192 384 768 1,152 1536
Price($) 4167 5,000 6700 11670 15300 20834 2250
Custom methylation chip(more than 480 samples)
Sites 96 methylation sites 384 methylation sites other
Price($) 41700 100000 inquriy

9.Methylation Analysis

a.Services

Bisulfite modification and sequencing method (BSP)

Frommer et al (1992) proposed methods of DNA methylation analysis, this method is reliable and precise. procedures:

  • Prepare genomic DNA of Sample
  • Bisulfite treatment (including purification and de-sulfosalicylic reaction) to convert unmethylated cytosine to uracil, while methylated cytosine remains unchanged.
  • Using methylation primer design software to design primers at the both ends of methylated regions.
  • Fragment amplification and PCR product recovery.
  • PCR products were sequenced directly or after cloned (quantitative analysis), analysis of each CpG island methylation.
  • Data Statistics. unmethylated DNA samples are used as a control.

Methylation specific PCR method (Methylation-Specific PCR, MS-PCR)

Herman et al (1996) in the use of bisulfite treatment on the basis of the new methods. High sensitivity of this method is mainly used for qualitative research. Procedures:

  • Prepare genomic DNA of Sample
  • Bisulfite treatment (including the purification and de-sulfosalicylic reaction).
  • Using methylation primer design software design methylation specific primers (primer 1) or non-methylated primers in methylated regions (primer 2) 2 pairs.
  • For specific PCR amplification, only the fragments fully combinate with methylated or non-methylation specific primers can amplify the product.
  • Detection of MSP amplification, if DNA methylation for the treatment chain primers amplified fragments, it shows that the detected methylation sites exist; if used for the treatment of non-methylated DNA chain The primers amplified fragments, then the detected methylation sites does not exist.

b.Fees

Services Quantity Unit price ($) Timeline Quantity Price
Bisulfite modification and sequencing method <=50 $110/sample/gene
(offer sequencing results of 5 clones )
15 to 20 business days >50 Inquire
$165/sample/gene
(offer sequencing results of 10 clones)
20 to 25 business days >50 Inquire
methylation specific PCR method <=100 $50/sample/gene 10 to 20 business days >100 Inquire
Combined Bisulfite Restriction Analysis <=100 $50/sample/site 15 to 20 business days >100 Inquire
High Resolution Melting method <=200 $45/sample/gene 15 to 20 business days >200 Inquire
Pyrosequencing <=200 $75/sample/gene 15 to 20 business days >200 Inquire
Methylation detection chip Sites of the amount of information Unit price /sample ($) The number of samples / chip Timeline
Human Methylation 450K 480000 $1,135 12 samples 4 weeks
Customized methylation chip 96 methylation sites Inquire 15 weeks
384 methylation sites Inquire 20 weeks

9.3 Results

Lab report (experimental steps, including the BSP method statistical results of 5 clones), PCR electrophoresis photographs, sequencing trace files.

10.Virus packaging

10.1 Adenovirus packaging services

10.2 Lentiviral packaging services

a. Service Quote

Services Quantity results Price($) Cycle
Vector construction ShRNA vector of the target gene 1 Report 166.7 1 to 2 weeks
Target gene shRNA vector Subscription 5 Report(4 object +1 negative contrast) 666.7 2 to 3 weeks
siRNA lentiviral screening Buy cell recovery and training 1 Cell state Image 83.5 2 working days
Slow virus infection experiments 1 The infection efficiency images and analysis 83.5 3 working days
shRNA design 3 Target information Free 1 working days
shRNA vector construction 4 Vector Construction Report 666.7 2 to 3 weeks
Lentiviral packaging
(10e8TU/ml)
5 The virus liquid and the titer test report 1333.5 2 to 3 weeks
The slower virus infection ( interference filter) 4 Picture 333.5 1 to 2 weeks
qRT-PCR primer design and verification 2 Primer sequences ( containing an internal reference ) 33.5 1 weeks
qRT-PCR detect 3*5 Experimental data and analysis 300 1 to 2 weeks
Western Blot detect 5 Experimental data and analysis ( primary antibody ) 333.5 1 to 2 weeks
Total price / / 3500 7 to 10 weeks
Establish stable Filter cell lines The effective siRNA lentiviral repeated infection ( positive + negative ) 2 Picture 266.7 1 to 2 weeks
Stable cell strain was amplified ( positive + negative ) 2 Cell picture 333.5 3 to 4 weeks
qRT-PCR detect 3*2 Experimental data and analysis 200 1 to 2 weeks
Western Blot detect 3*2 To provide an anti- experimental data analysis (customer ) 200 1 to 2 weeks
Infected cells (25T flasks , positive ( two kinds ) + negative , each bottle ) Cells and photo 333.5 1 to 2 weeks
Total price / / 1333.5 6 to 9 weeks

11.RNA Interference Service

a.Fees

Services Quantity Results Price ($) Timeline
shRNA vector construction Gene specific shRNA sequence is designed according to the target gene and cloned to shRNA expression vector 1 Report $165 1 to 2 weeks
Gene specific shRNA vector set 5 Report(4 gene specific shRNA vector +1 negative contrast) $665 2 to 3 weeks
siRNA lentiviral screening Cell culture 1 Image of cultured cells $85 2 business days
Lentiviral infection 1 Infection efficiency images and analysis $85 3 business days
shRNA design 3 Target information Free 1 business days
shRNA vector construction 4 Vector Construction Report $665 2 to 3 weeks
Lentiviral packaging
(10e8TU/ml)
5 The virus liquid and the titer test report $1330 2 to 3 weeks
The Lentiviral infection ( interference filter) 4 Picture $330 1 to 2 weeks
qRT-PCR primer design and verification 2 Primer sequences ( containing an internal reference ) $30 1 weeks
qRT-PCR detection 3*5 Experimental data and analysis $300 1 to 2 weeks
Western Blot detection 5 Experimental data and analysis ( primary antibody ) $330 1 to 2 weeks
Total price / / $3,500 7 to 10 weeks
Establish stable cell lines The effective siRNA lentiviral repeated infection ( positive + negative control) 2 Picture $265 1 to 2 weeks
Stable cell strain was amplified ( positive + negative control) 2 Cell picture $330 3 to 4 weeks
qRT-PCR detection 3*2 Experimental data and analysis $200 1 to 2 weeks
Western Blot detection 3*2 To provide an anti- experimental data analysis (customer ) $200 1 to 2 weeks
Infected cells (25T flasks , positive ( two kinds ) + negative , each bottle )   Cells and photo $330 1 to 2 weeks
Total price / / $1,330 6 to 9 weeks

b.Results

Reports include: siRNA sequences, construct vectors sequencing report, quantitative PCR test report, Western Blot image and all the experimental data and detailed experimental procedures