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Primer Synthesis Services

 

 

GENERAY Biotech Primer Synthesis Department has a GMP-grade primer synthesis laboratory and an industry-standard primer synthesis production line to provide high-quality products for scientific research users and the molecular diagnostics (IVD) industry. The company's leading products are Taqman probes, molecular beacons, double quenching probes, LNA probes, capture probes, DEL primers, NGS primers, RAA probes, RNA synthesis and other customized synthesis. The single synthesis specifications range from 2 OD to 10000 OD, and the synthesis length reaches 150base.


GENERAY Biological has five synthetic bases in Shanghai, Beijing, Guangzhou, Nanjing, and Hangzhou, providing faster and more considerate services for various users!

 

Common Primer:15-60bp

 

 

Low mutation rate: less than 1/1000


Provide 1~10 000OD specifications (can provide users with 96-well plate or 384-well plate package delivery)


Provide multiple purification methods including DSL, F-PAGE, HPLC and PAGE

 

Long-chain Primer: 61-90bp

 

 

Purity of polypropylene gel electrophoresis according to international standards


High-standard long-chain primers suitable for experimental requirements such as  NGS

 

Modified / Labeled (Probe) Services

 

 

GENERAY Biotech Primer Synthesis provides multiple types of modified / labeled primers, including: fluorescent probe synthesis services, capture probe synthesis services, fluorescent in situ hybridization services, qPCR probe design and synthesis services. Available modification types are: FAM, HEX, TET, TAMRA, ROX, VIC, NED, Alexa Flour, ATTO series, CY series, etc., amino modification, thiol modification, biotin, electrochemical labeling, Fluorescence, BHQ0, BHQ1, BHQ2, BHQ3, Dabcyl, ECL, NFQ, FAM-dT, BHQ-dT, dSpacer et.al.

 

Base Modification

 

 

Modification

Purification way

Phosphorothioate (S-oligos)*

PAGE/HPLC

Deoxylnosine (dI)

PAGE/HPLC

DeoxyUridine (dU)

PAGE/HPLC

5-Methyl dC

PAGE/HPLC

2'-OMe- C

PAGE/HPLC

2'-OMe- G

PAGE/HPLC

2'-OMe- T

PAGE/HPLC

2'-OMe- A

PAGE/HPLC

5-Carboxy-Dc (5Cac)

PAGE/HPLC

5-Hydroxymethyl-dC (5hmc)

PAGE/HPLC

5-Formyl-dC (5fmc)

PAGE/HPLC

N6-Me-dA

PAGE/HPLC

2’-F-base

PAGE/HPLC

LNA-dA/dG/dC/dT

PAGE/HPLC

Note: HPLC purification method is charged separately

 

Single Labeled Probe- Fluorescent Modification

 

Modification

5’ modified

Middle modified

3’ modified

FAM

FITC

HEX

SIMA

×

JOE

ROX

Rhodamine Green

Rhodamine Red

TAMRA

TET

Texas Red

Cy3

Cy5

AMCA

BHQ -1

×

BHQ -2

×

Eclipse

×

×

Dabsyl

×

Fluorescein

ATTO550

ATTO565

ATTO633

Yakima Yellow

×

iFluor™ 488

iFluor™ 532

iFluor™ 555

iFluor™ 648

iFluor™ 750

 

Double Probe Modification

 

5’-Fluorophore

3'-Quencher

 

 

5'- FAM

3'- TAMRA

3'- Dabcyl

3'- BHQ1

3'-Eclipse

3'-MGB

 

 

5'- HEX

3' TAMRA

3'- Dabcyl

3'- BHQ1

3'- BHQ2

3'-Eclipse

3'-MGB

5'-TET

3'- Dabcyl

3'- TAMRA

3'- BHQ1

3'- BHQ2

3'-Eclipse

5'-JOE

3'- TAMRA

3'- Dabcyl

3'- BHQ2

5'-ROX

3'- BHQ2

3'-Eclipse

3'-MGB

5'-Cy3

3'- Dabcyl

3'-Eclipse

3'- BHQ2

5'-Cy5

3'- Dabcyl

3'-Eclipse

3'- BHQ3

3'-MGB

5'- TAMRA

3'- BHQ2

3'-Eclipse

5’-VIC

3'-MGB

5’-NED

3'-MGB

 

Chemical Modification

 

Modification

5’ modified

Middle modified

3’ modified

Phosphorylation

Aminolinker

Spacers

Thiolation

Biotin

Biotin TEG

×

Digoxigenin

 

Connection Probe

 

Modification

Purification way

Triple SH

HPLC

Maleimide

HPLC

Azide(N3)

HPLC

CHCH

HPLC

DBCO

HPLC

COOH

HPLC

Acryl

HPLC

 

Please call for other Modifications

 

More Modification

VIC

Alexa fluor 488

N6-Me-dA

AMCA

MAX

Alexa Fluor 532

Azobenzene

Acrydite

NED

Alexa fluor 546

Bromo dU

CY3.5

PET

Alexa fluor 555

1-Me-dA

CY5.5

TAZ

Alexa fluor 568

PC-Linker

CY7.5

SID

Alexa fluor 633

Inverted base

QSY

LIZ

Alexa fluor 647

Pyrene

APhluor 593

Qusar570

Alexa fluor 750

Symmetric

            IRDye ®

Qusar670

Bodipy 530/550

CHO

5' Terminal Caps

 

Commonly used Fluorescent Dye Parameters

 

Dye

Excitation(nm)

Emission(nm)

Colour

6-FAM

494

518

Yellow-Green

Flurescein (FITC)

495

520

TET

521

536

JOE

520

548

Yellow

HEX

533

558

CY3

550

570

Yellow-Orange

TAMRA

544

576

ROX

576

601

Orange

CY5

650

670

Red

 

Synthesis Features

 

Advanced synthesis equipment: high-throughput Dr.Oligo series and large-scale Mermade series synthesizers


Various purification methods: DSL, F-PAGE, HPLC and PAGE


Good product quality: relying on ESI-MS mass spectrometer and HPLC chromatography for quality control


Fast synthesis speed: Set up synthesis bases in Shanghai, Beijing, Guangzhou (Kunming), Nanjing, and Hangzhou to fully guarantee the synthesis speed

 

Product Quality

 

Correctness: Strict quality control standards and QC methods ensure product correctness


Accuracy: Retest the concentration of each probe and compare with the standard, accurately control the difference between batches, and ensure the accuracy of the product


Stability: using unique technology to reduce the damage of fluorescein during purification


Quality control: perfect central control, real-time monitoring of the coupling efficiency of each base, to ensure that the efficiency of each step is greater than 99%; ESI-MS mass spectrometry, HPLC, full-wavelength scanning, and fluorescence spectrum detection PAGE analysis gel detection and CE capillary electrophoresis detection 6-fold QC

 

Primer Purification Guide

 

Primer
length

Purification Way

DSL

F-PAGE

PAGE

HPLC

<15nt

Applicable

Not Applicable

Not Applicable

Recommend

15-40nt

Recommend

Recommend

Applicable

Recommend

41-59nt

Applicable

Applicable

Recommend

Applicable

60-120nt

Not Applicable

Not Applicable

Recommend

Applicable(60-100nt)

Scope of application

PCR amplification, whole gene synthesis, DNA sequencing

DNA sequencing, whole gene synthesis, site-directed mutation, PCR cloning, quantitative PCR, multiplex PCR, point mutation / subcloning, RNA interference / gene construction

Purity requirements for next-generation sequencing, diagnostic PCR, mutation library construction, and protein-binding gel migration electrophoresis analysis

Dot cloning, subcloning, diagnostic PCR and protein binding gel migration electrophoresis analysis; modified primers with hydrophobic groups; commercial diagnostic primers and probes

 

DNA Production Quality Control Flow Chart

 

DNA Production Quality Control Report-HPLC Primer purity test

 

Explanation of test results: In the figure, the area of the peak accounts for 100% . So the test is qualified.
Eligibility criteria: The main peak area is 90% or more.

 

DNA Production Quality Control Report-MS

  

MS test result description: Observing the theoretical molecular weight and actual molecular weight, it can be found that the theoretical molecular weight of this primer are 7464.065537.86、9357.35、 7820.35 and 8190.07, and the actual molecular weight are 7465.05538.7、9358.67822.4 and 8187.2.  The test is qualified.

Eligibility criteria: The measured molecular weight and the molecular weight less than 0.1% difference theory

 

Have a Question? 

 

Order Online:   dna@generay.com.cn

 

By Phone:    86-21-67840801/67840802