Molecular Cloning And Sub Cloning

iLife offers following services for cloning:

1. Cloning and Sequencing of PCR Products in our standard vector that would enable the scientists to acquire complete and analyzed sequence data of their PCR products.

Part #	Different Strategies
GCL001	Sub-Cloning of DNA of < 1 kb with Primers provided by the researcher
GCL002	Sub-Cloning of DNA of 1 kb with Primers provided by the researcher
GCL003	Sub-Cloning of DNA of 2 kb with Primers provided by the researcher
GCL004	Sub-Cloning of DNA of 3 kb with Primers provided by the researcher
GCL005	Sub-Cloning DNA of < 1 kb including Primer designing and synthesis
GCL006	Sub-Cloning DNA of 1 kb including Primer designing and synthesis
GCL007	Sub-Cloning DNA of 2 kb including Primer designing and synthesis
GCL008	Sub-Cloning DNA of 3 kb including Primer designing and synthesis

Acceptable:

• Cells,
• gDNA,
• RNA
• Single stranded cDNA (gene specific primer is required)
• Double stranded cDNA 
• Plasmid (in case of subcloning) and
• Gel purified PCR purified supported by gel image

Precautions and limitations:
If PCR product is provided:

•  The band of interest should be the most prominent product within the PCR sample.
• The samples should not have excessive smearing.
• Please indicate if the PCR product has a higher GC – content

Deliverables:

• Clone DNA (up to 5 μg)
• Sequence data (bi-directional, raw data)
• Final analyzed sequence data
• Project Report

Please Enquire for More Information

 

2. Cloning of DNA for Expression of Protein

iLife can assist in cloning any gene in any vector for expression of the protein product. Based on the requirement, the derived protein can have:

• His Tag
• Maltose binding protein domain
• FLAG

Part #	Description
GCL009	Sub-Cloning of DNA of < 1 kb for Expression of Proteins w/Primers from Researcher
GCL010	Sub-Cloning of DNA of 1 kb for Expression of Proteins w/Primers from Researcher
GCL011	Sub-Cloning of DNA of 2 kb for Expression of Proteins w/Primers from Researcher
GCL012	Sub-Cloning of DNA of 3 kb for Expression of Proteins with Primers Provided
GCL013	Sub-Cloning DNA of < 1 kb for Expression of Proteins including Primer Designing and Synthesis
GCL014	Cloning DNA of 1 kb for Expression of Proteins including Primer Designing and Synthesis
GCL015	Cloning DNA of 2 kb for Expression of Proteins including Primer Designing and Synthesis
GCL016	Cloning DNA of 3 kb for Expression of Proteins including Primer Designing and Synthesis

Acceptable:

• Cells,
• gDNA,
• RNA
• Single stranded cDNA (gene specific primer is required)
• Double stranded cDNA 
• Plasmid (in case of subcloning) and
• Gel purified PCR purified supported by gel image

Precautions and limitations:

• If PCR product is provided:
•  The band of interest should be the most prominent product within the PCR sample.
• The samples should not have excessive smearing.
• Please indicate whether the PCR product has a higher GC – content

Deliverables:

• Clone DNA (up to 5 μg)
• Sequence data (bi-directional, raw data)
• Final analyzed sequence data
• Project Report

Please Enquire for More Information

 

3. Cloning and Sequencing of Up-stream/ Down-stream Regions from Known Sequence

iLife can assist in the cloning and sequencing of the Regulatory Elements of any gene, which includes  promoter, enhancer or 5’-untranslated region in up-stream sequences and 3’-untranslated region, terminator sequences etc. at down-stream sequences,  are required for gene regulation, expression and m-RNA stability studies.

This service will include:

• Cloning of up to 2 kb of up-stream or downstream sequences from known sequence data
• Confirmed bi-directional sequence data
• Putative regulatory regions indicated

Part #	Description
GCL017	Cloning and Sequencing of Up-stream/ Down-stream Regions from Known Sequence with Primers provided by Researcher
GCL018	Cloning and Sequencing of Up-stream/ Down-stream Regions from Known Sequence including Primer designing and synthesis

Deliverables:

• Cloned DNA containing up to 2 kb of up-stream/down-stream sequence
• Bi-directional sequence data (raw data as well as analyzed and aligned data) with the putative regulatory regions indicated
• Project report

Please Enquire for More Information

 

4. Cloning of STRs/ Microsatellite Markers

This service will include:

• Development of 10 independent STR clones, starting from samples or genomic DNA (minimum guaranteed)
• Sequencing of the clones, demonstrating STRs
• STR specific primers would be designed (optional)
• Standardization of STR specific “PCR/multiplex PCR” conditions as well as screening samples for polymorphism are also undertaken (optional)

Part #	Description
GCL019	Cloning of STRs/ Microsatellite Markers with Primers provided
GCL020	Cloning of STRs/ Microsatellite Markers including Primer designing and synthesis

Deliverables:

• 10 clones with STRs/ minimum
• Sequence data and electropherogram
• Final report

Please Enquire for More Information

 

5. Full-Length cDNA Cloning Services

Cloning and sequencing of full-length cDNA allows scientists to:

• Analyse the intron/ exon junctions;
• Deduce the amino acid sequence coded by the gene;
• Study the up-stream/ down-stream regulatory regions;
• Express the gene for protein expression studies etc.

Either of the approach can be implemented to for cloning of full-length cDNA:

• Partial/ full-length cDNA sequence data
• Genomic clone sequence data
• Amino acid sequence data
• cDNA/ genomic DNA libraries

This service will include:

• Cloning into any commercially available cloning and/ or expression vector
• Cloning and sequencing of full-length cDNA irrespective of the gene size and the number of introns present.
• Fast and reliable service

 

Part #	Description
GCL021	Cloning of Full-length cDNA of 1 kb with Primers provided by researcher
GCL022	Cloning of Full-length cDNA of 2 kb with Primers provided by researcher
GCL023	Cloning of Full-length cDNA of 3 kb with Primers provided by researcher
GCL024	Cloning of Full-length cDNA of 1 kb including Primer designing and synthesis
GCL025	Cloning of Full-length cDNA of 2 kb including Primer designing and synthesis
GCL026	Cloning of Full-length cDNA of 3 kb including Primer designing and synthesis

Deliverables:

• Full-length cDNA into a cloning vector
• Bi-directional sequence data (both raw data and analysed data)
• Project report

Please Enquire for More Information

 

6. Customized cloning services will include designing and synthesis of primers, PCR, restriction digestion and ligation with the vector.

Please Enquire for More Information