In the experiments of expression of target protein and construction of expression vector, it is usually necessary to obtain a segment of target gene. According to the known reference sequence (published on NCBI) provided by you, we can use genomic DNA or total RNA as template to obtain a gene in a region you need.
1.Total RNA extraction.
2.RNA extraction (if necessary).
3.Reverse transcription synthesis of the first strand of DNA and amplification of double-stranded DNA by PCR.
4.Subcloning to other expression vectors (if necessary) generally takes 2-4 weeks depending on the difficulty of the experiment.
|experimental item/fragment length||less than 1000bp||1000-2000bp||2000-3000bp|
|total RNA extraction||inquiry||inquiry||inquiry|
|RNA extraction (alternative)||inquiry||inquiry||inquiry|
|reverse transcription synthesis of the first strand of DNA and amplification of double-stranded DNA by PCR||inquiry||inquiry||inquiry|
|purification of PCR products, cloning of PCR products to standard vector pUC57-T and sequencing (usually only one clone, if customers need to sequence multiple clones, additional sequencing costs)||inquiry||inquiry||inquiry|
|subcloning to other expression vectors (alternative)||inquiry||inquiry||inquiry|
1.The samples we currently receive are in the form of materials identical to the reference sequence species or genomic DNA/total RNA or fresh tissues or cells of the materials mentioned above.
The experimental material must be fresh. Please put it into liquid nitrogen immediately after sampling or add sample stabilizer (such as Sample Protector).
2.Genomic DNA/total RNA did not degrade significantly, the total amount was more than 5 mg. If the gene abundance is low or the selected gene is long, in order to ensure the smooth experiment, please provide as many materials as possible.
3.Because of the individual differences of materials and mutations in the process of PCR, we do not guarantee that the obtained genes are identical with the base of the reference sequence.
2.sequencing color picture
3.digestion site picture
4.plasmids containing target genes and puncture bacteria
ORF CDNA Clones Service
TA Cloning And Sequencing Service
Gene Synthesis Services