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Ready to use CRISPR Cas9 sgRNA Synthesis .pdf



Original filename: Ready-to-use CRISPR Cas9 sgRNA Synthesis.pdf
Title: Ready-to-use CRISPR Cas9 sgRNA Synthesis
Author: http://www.synbio-tech.com/services/crispr-cas9/crispr-cas9-sgrna-design-center/grna-synthesis/

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Ready-to-use CRISPR Cas9 sgRNA Synthesis
CRISPR-Cas9 is one of the most convenient types of gene editing technology, and
has been widely used in editing genes of many species[1]. CRISPR-Cas9 generally
involves construction a plasmid containing the specific endogenous promoter and
connection of the Cas9 gene and a synthetic sgRNA, followed by transfer into the
cell or animal. However, plasmid construction and verification are both tedious and
time-consuming. The degradation of the plasmid is relatively slow, which may prove
to be inconvenient for subsequent experiments.
To improve the efficiency of the CRISPR-Cas9 system, Synbio Technologies has
developed ready-to-use sgRNA synthesis services. We can perform sgRNA target
design, DNA template synthesis of sgRNA, sgRNA in vitro transcription, and sgRNA
purification, to provide customers ready-to-use sgRNA that can be directly
transfected into cells or animals. Synbio Technologies’s ready-to-use sgRNA saves
time on plasmid construction, and avoids the drawbacks of potentially
non-degraded plasmids.
According to the publications, in vitro transcription of sgRNA has successfully edited
the genes of different species including zebrafish[2], mouse[3]as well as filamentous
fungi[4]etc. Synbio Technologies has also designed 3 universal negative control
sgRNA for Human and Rat genome: Syno®-negative controls sgRNA1, sgRNA2, and
sgRNA3. These sequences are used as a negative control in Human and Rat
gene/genome editing experiments.

Service Advantages:
 One-stop solution: Synbio Technologies provides integrated services from
sgRNA target design to high purity ready-to-use sgRNA production.
 Fast delivery: In just 3 business days, Synbio will deliver up to 20ug of
customized ready-to-use sgRNA
 Convenience: ready-to-use sgRNA can be directly inject into animals or
transfected into cells, improving the efficiency of gene editing experiments

Case Study – Synbio Technologies:
Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com

Synbio Technologies has designed 8 sgRNAs targeting several genes in mouse, and
performed in vitro transfection. The experimental period was shortened to 2 days,
and the sgRNA amount was increased to 10-20 ug. This change could mean a
significant jump in efficiency for synthetic biology experiments utilizing
CRISPR-Cas9.

Workflow:

One-step synthesis of DNA template

Result:
 Clone DNA template into pUC57 vector, the sequencing result (Fig. 1) coincided
with the designed sequence.

Fig. 1. Comparison between blunt end ligation result and designed sequence

 Agarose gel electrophoresis of sgRNA obtained by in vitro transcription, clear
bands shown in Fig. 2.
Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com

Fig. 2. agarose gel electrophoresis of sgRNA
 sgRNA verification: Transcript sgRNA into cDNA, design sgRNA amplification
primer, and obtain the complementary DNA sequence by PCR reaction. Clone
DNA sequence into pUC57 vector; sequencing result (Fig. 3) showed the sgRNA
sequence is correct.

Fig. 3. sgRNA sequence verification using agarose gel electrophoresis
*The template of Lane 1 is reverse transcripted cDNA, The template of Lane 2 is sgRNA
digested by DNaseI; The template of Lane 3 is in vitro transcripted DNA

Service Specifications:

Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com

Turnaround
Product/Service
Time
Service Name
Specifications
(business
day)

Deliverables

Price

sgRNA design
<10 sgRNA, 5
DNA template
Ready-to-use
10-20 sgRNA,
synthesis
sgRNA
10
In vitro sgRNA
synthesis
>20 sgRNA,I
transcription and
nquiry
purification

sgRNA
COA

Inquiry

Negative control
sgRNA
Syno® negative
In vitro sgRNA
control sgRNA
transcription and
purification

5days

Syno®-negative control
sgRNA1
Syno®-negative control
Inquiry
sgRNA2
Syno®-negative control
sgRNA3

Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com


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