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Long single-stranded DNA (ssDNA) can be easily prepared Long ssDNA Preparation Kit for 1.5kb & 3.0kb

Date：August 13 2019Web Page No：81351

BioDynamics Laboratory Inc

This kit can prepare long single-stranded DNA (≦1.5 kb or 1.5-3 kb) with high purity and accurate sequence by nicking enzyme amplification reaction.

☞ 3-10 kb ssDNA preparation kit is also available. Please check it.

High-purity long single-stranded DNA with the target sequence can be easily obtained.

Since the preparation of long single-stranded DNA (ssDNA) involves steps such as PCR, use of synthetic DNA oligomers, exonuclease reaction, and reverse transcriptase reaction, it was inevitable that internal mutations and terminal deletions would occur. In addition, denaturing acrylamide gel electrophoresis and streptavidin-coated paramagnetic beads have been used for purification, then there have been cases in which large amounts of double-stranded DNA (dsDNA) have been detected.
With the Long ssDNA Preparation Kit (this product), you can obtain long ssDNA with the sequence of interest in high purity with simple operation.

Features

  • Nicking endonucleases allows the preparation of long ssDNA without mutations or terminal deletions.
  • The entire procedure is similar to that of conventional dsDNA fragments, and no special equipment or reagents are required.
  • No PCR amplification, no errors
  • By using with Denaturing Gel-loading Buffer, the long ssDNA of interest can be separated and purified by agarose gel electrophoresis.
  • The Long ssDNA Gel Extraction Kit (#DS640) is also included.

Useful for genome editing
By using the long ssDNA prepared with this kit as donor DNA and introducing it together with CRISPR/Cas9, ☞ we succeeded in accurately and efficiently knocking in the GFP sequence.


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Plasmid Map

for ≦1.5 kb for 1.5-3 kb
Plasmid Map for preparing long ssDNA, DS615 Plasmid Map for preparing long ssDNA, DS625
☞ Vector sequence for DS615 ☞ Vector sequence for DS625

※ The above sequence file has a password (Access ID). Please enter the Access ID shown on the box. If you have previously purchased and cannot find the Access ID, please contact us.


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Principle & Procedure

Principle & Procedure for long ssDNA preparation kit

  1. The DNA of interest is cloned into a plasmid using a pair of two nicking endonuclease sites or a combination of a nicking endonuclease site and a restriction enzyme site.
  2. The resulting plasmid harboring the DNA is digested with a pair of two nicking endonucleases or a combination of a nicking endonuclease and a restriction enzyme.
  3. The nicked plasmid is denatured and then subjected to agarose gel electrophoresis.
  4. The band corresponding to a long ssDNA is excised and extracted.

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Application Example

Usage case for long ssDNA preparation kit
  • Lane 1: pLSODN-1 transfected with DNA fragment (1.5 kb) and treated with Nicking enzyme
  • Lane 2: Purified long ssDNA (1.5 kb)
  • Lane 3: pLSODN-3 transfected with DNA fragment (3.0 kb) and treated with Nicking enzyme
  • Lane 4: Purified long ssDNA (3.0 kb)

Long ssDNA (1.5 kb & 3.0 kb) prepared by Long ssDNA Preparation Kit (LsODN Preparation Kit)

The target dsDNA fragment (1.5 kb or 3.0 kb) was cloned into the MCS (between Nt.BspQI sute and Nb.BsrDI site) of pLSODN-1 or pLSODN -3. This fragment was treated with nicking enzyme (Nt.BspQI and Nb.BsrDI) to introduce nicks at both ends of the desired fragment. After denaturation, the samples were separated by agarose gel electrophoresis, and the DNA from the target band was extracted and purified. All samples were denatured by denaturing Gel-loading buffer and electrophoresed.


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Kit Contents

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Citation

  1. "ssODN-mediated knock-in with CRISPR-Cas for large genomic regions in zygotes."
    Yoshimi K. et al., Nat. Commun., 7, 10431 (2016). Nature Communications. 20;7:10431.
    Long ssDNA, poly(A)-loaded Cas9-poly(A) RNA, and guide RNA (Thy1-TGA) prepared by nicking endonuclease (this product) were microinjected into fertilized rat eggs, and the entire GFP gene was successfully knocked into the C-terminus of the rat rThy1 gene with high efficiency (11.1%).
    This paper shows that the knockin efficiency of the entire length of a foreign gene can be increased by the LssDNA method using long single-stranded DNA (LssDNA vs ssODN).
  2. "Long noncoding RNA lncKdm2b is required for ILC3 maintenance by initiation of Zfp292 expression."
    Liu B., et al., Nat. Immunol., 18 (5), 499 (2017). [PMID:28319097]
  3. "The ER membrane adaptor ERAdP senses the bacterial second messenger c-di-AMP and initiates anti-bacterial immunity."
    Xia P., et al., Nat. Immunol., 19 (2), 141 (2018). [PMID:29292386]
  4. "LncGata6 maintains stemness of intestinal stem cells and promotes intestinal tumorigenesis."
    Zhu P., et al., Nat. Cell. Biol., 20 (10), 1134 (2018). [PMID:30224759]
  5. "Long noncoding RNA lncHand2 promotes liver repopulation via c-Met signaling."
    Wang Y., et al., J Hepatol., 69 (4), 861 (2018). [PMID:29653123]
  6. "IL-13 secreted by ILC2s promotes the self-renewal of intestinal stem cells through circular RNAcircPan3."
    Zhu P., et al., Nat. Immunol., 20 (2), 183 (2019). [PMID:30643264]
  7. "Exonuclease III (XthA) Enforces InVivo DNA Cloning of Escherichia coli To Create CohesiveEnds."
    Nozaki S. & Niki H., J. Bacteriol., 201 (5), e00660-18 (2019). [PMID:30530516]
  8. "New Additions to the CRISPR Toolbox: CRISPR- CLONInG and CRISPR- CLIP for Donor Construction in Genome Editing."
    Shola D.T.N., et al., CRISPR J., 3 (2), 109 (2020). [PMID:32315232]
  9. "Efficient CRISPR-Cas9-Mediated Knock-In of Composite Tags in Zebrafish Using Long ssDNA as a Donor."
    Ranawakage D.C., et al., Front. Cell. Dev. Biol., 8, 598634 (2021). [PMID:33681181]
  10. "Oocyte-specific gene knockdown by intronic artificial microRNAs driven by Zp3 transcription in mice."
    Sasaki K., et al., J. Reprod. Dev., 67 (3), 229 (2021). [PMID:33716236]
  11. "Hepatitis B virus cccDNA is formed through distinct repair processes of each strand."
    Wei L., et al., Nat. Commun., 12 (1), 1591 (2021). [PMID:33707452]

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Precautions for purchase

  • Letter of Assurance (LoA) is required upon ordering.
  • If vector products will be used for commercial purposes (e.g. creating genetically modified models or offering contract services to any other party by using vector products), license is required separately. Please contact us in advance.
  • The price is different for academic and commercial entities.

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Product Information

for ≦ 1.5 kb ssDNA preparation kit

[Date : April 25 2025 00:06]

Detail Product Name Product Code Supplier Size Price
Long ssDNA Preparation Kit for 1.5kb, Academic
DatasheetThis may not be the latest data sheet. 		DS615 BDLBioDynamics Laboratory Inc 1 kit $800

Description The kit provides a simple and easy method for generation of a long ssDNA (within 1.5 kb). Letter of Assurance (LoA) is required for purchase. Please ask your local distributor or Funakoshi. For commercial purpose use, license contract should be made between customers and Funakoshi. Please ask Funakoshi for detail.
Storage RT,-20°C CAS
Link

Long ssDNA Preparation Kit for 1.5kb, Commercial Entities
DatasheetThis may not be the latest data sheet. 		DS615 BDLBioDynamics Laboratory Inc 1 kit $1,600

Description The kit provides a simple and easy method for generation of a long ssDNA (within 1.5 kb). Letter of Assurance (LoA) is required for purchase. Please ask your local distributor or Funakoshi. For commercial purpose use, license contract should be made between customers and Funakoshi. Please ask Funakoshi for detail.
Storage RT,-20°C CAS
Link

[Date : April 25 2025 00:06]

Long ssDNA Preparation Kit for 1.5kb, Academic

DatasheetThis may not be the latest data sheet.


  • Product Code: DS615
  • Supplier: BDL
  • Size: 1kit
  • Price: $800

Description The kit provides a simple and easy method for generation of a long ssDNA (within 1.5 kb). Letter of Assurance (LoA) is required for purchase. Please ask your local distributor or Funakoshi. For commercial purpose use, license contract should be made between customers and Funakoshi. Please ask Funakoshi for detail.
Storage RT,-20°C CAS
Link

Long ssDNA Preparation Kit for 1.5kb, Commercial Entities

DatasheetThis may not be the latest data sheet.


  • Product Code: DS615
  • Supplier: BDL
  • Size: 1kit
  • Price: $1,600

Description The kit provides a simple and easy method for generation of a long ssDNA (within 1.5 kb). Letter of Assurance (LoA) is required for purchase. Please ask your local distributor or Funakoshi. For commercial purpose use, license contract should be made between customers and Funakoshi. Please ask Funakoshi for detail.
Storage RT,-20°C CAS
Link

for 1.5-3 kb ssDNA preparation kit

[Date : April 25 2025 00:06]

Detail Product Name Product Code Supplier Size Price
Long ssDNA Preparation Kit for 3kb, Academic
DatasheetThis may not be the latest data sheet. 		DS625 BDLBioDynamics Laboratory Inc 1 kit $800

Description The kit provides a simple and easy method for generation of a long ssDNA (within 1.5-3 kb). Letter of Assurance (LoA) is required for purchase. Please ask your local distributor or Funakoshi. For commercial purpose use, license contract should be made between customers and Funakoshi. Please ask Funakoshi for detail.
Storage RT,-20°C CAS
Link

Long ssDNA Preparation Kit for 3kb, Commercial Entities
DatasheetThis may not be the latest data sheet. 		DS625 BDLBioDynamics Laboratory Inc 1 kit $1,600

Description The kit provides a simple and easy method for generation of a long ssDNA (within 1.5-3 kb). Letter of Assurance (LoA) is required for purchase. Please ask your local distributor or Funakoshi. For commercial purpose use, license contract should be made between customers and Funakoshi. Please ask Funakoshi for detail.
Storage RT,-20°C CAS
Link

[Date : April 25 2025 00:06]

Long ssDNA Preparation Kit for 3kb, Academic

DatasheetThis may not be the latest data sheet.


  • Product Code: DS625
  • Supplier: BDL
  • Size: 1kit
  • Price: $800

Description The kit provides a simple and easy method for generation of a long ssDNA (within 1.5-3 kb). Letter of Assurance (LoA) is required for purchase. Please ask your local distributor or Funakoshi. For commercial purpose use, license contract should be made between customers and Funakoshi. Please ask Funakoshi for detail.
Storage RT,-20°C CAS
Link

Long ssDNA Preparation Kit for 3kb, Commercial Entities

DatasheetThis may not be the latest data sheet.


  • Product Code: DS625
  • Supplier: BDL
  • Size: 1kit
  • Price: $1,600

Description The kit provides a simple and easy method for generation of a long ssDNA (within 1.5-3 kb). Letter of Assurance (LoA) is required for purchase. Please ask your local distributor or Funakoshi. For commercial purpose use, license contract should be made between customers and Funakoshi. Please ask Funakoshi for detail.
Storage RT,-20°C CAS
Link



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Kit Component

Gel Extraction Kit

Spin column kit for extraction of ssDNA from gel.

[Date : April 25 2025 00:06]

Detail Product Name Product Code Supplier Size Price
Long ssDNA Gel Extraction Kit for 3kb
DatasheetThis may not be the latest data sheet. 		DS640 BDLBioDynamics Laboratory Inc 25 tests $300

Description Long ssDNA Gel Extraction Kit for 1.5 - 3kb.
Storage RT CAS
Link

[Date : April 25 2025 00:06]

Long ssDNA Gel Extraction Kit for 3kb

DatasheetThis may not be the latest data sheet.


  • Product Code: DS640
  • Supplier: BDL
  • Size: 25tests
  • Price: $300

Description Long ssDNA Gel Extraction Kit for 1.5 - 3kb.
Storage RT CAS
Link


Denaturing Gel-Loading Buffer

[Date : April 25 2025 00:06]

Detail Product Name Product Code Supplier Size Price
Denaturing Gel-Loading Buffer
DatasheetThis may not be the latest data sheet. 		DS612 BDLBioDynamics Laboratory Inc 2x1 ml $150

Description
Storage -20°C CAS
Link

Long ssDNA Preparation Kit for 10kb

Denaturing Gel-Loading Buffer
DatasheetThis may not be the latest data sheet. 		DS611 BDLBioDynamics Laboratory Inc 5x1 ml $300

Description
Storage -20°C CAS
Link

Long ssDNA Preparation Kit for 10kb

[Date : April 25 2025 00:06]

Denaturing Gel-Loading Buffer

DatasheetThis may not be the latest data sheet.


  • Product Code: DS612
  • Supplier: BDL
  • Size: 2x1ml
  • Price: $150

Description
Storage -20°C CAS
Link

Long ssDNA Preparation Kit for 10kb

Denaturing Gel-Loading Buffer

DatasheetThis may not be the latest data sheet.


  • Product Code: DS611
  • Supplier: BDL
  • Size: 5x1ml
  • Price: $300

Description
Storage -20°C CAS
Link

Long ssDNA Preparation Kit for 10kb



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Related Product

for 3-10 kb ssDNA preparation kit

Challenge the limits of long single-stranded DNA! ssDNA up to 10 kb can be prepared.

☞ Learn more about the Long ssDNA Preparation Kit for 10 kb


Molecular weight ladder marker

[Date : April 25 2025 00:06]

Detail Product Name Product Code Supplier Size Price
Dynamarker, Prestain Marker for RNA high
DatasheetThis may not be the latest data sheet. 		DM260 BDLBioDynamics Laboratory Inc 180 µl $260

Description Prestain RNA ladder marker. Approximately 30 loadings.
Storage -80°C CAS
Link

Molecular Biology Product List
Prestained RNA Ladder

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DatasheetThis may not be the latest data sheet. 		DM122 BDLBioDynamics Laboratory Inc 1 set $150

Description DynaMarker DNA High D is useful for determination of DNA fragments 300 bp to 10,000 bp in size. 100tests. Supplied with 6XBPB loading dye(1ml).
Storage -20°C CAS
Link

Molecular Biology Product List

[Date : April 25 2025 00:06]

Dynamarker, Prestain Marker for RNA high

DatasheetThis may not be the latest data sheet.


  • Product Code: DM260
  • Supplier: BDL
  • Size: 180µl
  • Price: $260

Description Prestain RNA ladder marker. Approximately 30 loadings.
Storage -80°C CAS
Link

Molecular Biology Product List
Prestained RNA Ladder

DynaMarker, DNA High D

DatasheetThis may not be the latest data sheet.


  • Product Code: DM122
  • Supplier: BDL
  • Size: 1set
  • Price: $150

Description DynaMarker DNA High D is useful for determination of DNA fragments 300 bp to 10,000 bp in size. 100tests. Supplied with 6XBPB loading dye(1ml).
Storage -20°C CAS
Link

Molecular Biology Product List

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