cDNA:
PCR Ready First Strand cDNA is an excellent source of tissue specific, PCR-ready cDNA, and it can be immediately used for gene discovery or expression analysis. First-Strand cDNA is synthesized from RNA isolated from a wide variety of documented human adult and fetal normal tissues, human diseased and tumor tissues, mouse, rat, monkey and plant tissues. Total RNA used for cDNA synthesis is isolated by modified guanidine thiocyanate techniques.

A sampling of Restriction Endonucleases:

EcoRI
5'-G^A A T T C-3'
3'-C T T A A^G-5'
EcoRI cuts neither Gm6AATTC nor GAm6ATTC, but cuts GAATTm5C, GAATThm5C and GAAhm5Uhm5UC.

BamHI
5'-G^G A T C C-3'
3'-C C T A G^G-5'
BamHI does not cut GGATm4CC, GGATm5CC, GGAThm5Chm5C, GGAhm5UCC, but cuts GGATCm5C, GGm6ATCC, GGm6ATCm5C and GGATCm4C

PstI
5'-C T G C A^G-3'
3'-G^A C G T C-5'
Dam/Dcm/CpG/EcpKl/EcoBl: never overlaps- no effect

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Cloning :
Did you know United States Biological has a wide range of products geared for your cloning lab? We carry plasmids like pUC18 and pUC19, oligos, cDNA probes, polymerases, reverse transcriptases as well as staples like agaroses and XGal.

A variety of specialized kits are also available.

Complementary DNA (cDNA) is DNA synthesized from a mature mRNA template in a reaction catalyzed by the enzyme reverse transcriptase. This enzyme operates on a single strand of mRNA, generating its complementary DNA based on the pairing of RNA base pairs (A, U, G and C) to their DNA complements (T, A, C and G respectively).

Often it is desirable to express eukaryotic genes in prokaryotic cells (e.g., to produce recombinant proteins in large quantities).  A simplified method of doing this involves the addition of eukaryotic DNA to a prokaryotic host, which transcribes the cDNA to mRNA and then translates it to protein.
One difference between eukaryotic and prokaryotic genes is that eukaryotic genes can contain introns, which are not coding sequences, and must be spliced out of the RNA primary transcript before it becomes mRNA and can be translated into protein. Prokaryotic genes have no introns, so their RNA is not subject to splicing

To obtain eukaryotic cDNA whose introns have been spliced: A eukaryotic cell transcribes the DNA (from genes) into RNA (pre-mRNA). The same cell processes the pre-mRNA strands by splicing out introns, and adding a poly-A tail and 5’ Methyl-Guanine cap. This mixture of mature mRNA strands is extracted from the cell. A poly-T oligonucleotide primer is hybridized onto the poly-A tail of the mature mRNA template. Reverse transcriptase requires this double-stranded segment as a primer to start its operation. The reverse transcriptase scans the mature mRNA and synthesizes a sequence of DNA that complements the mRNA template, hence the name cDNA.

USBio’s first strand cDNA is prepared by using MMLV Reverse Transcriptase.  MMLV reverse transcriptase (encoded by Moloney Murine Leukemia Virus) is an RNA-dependent DNA polymerase that synthesizes the complementary cDNA first strand from a single-stranded RNA template to which a primer has been hybridized. MMLV reverse transcriptase will also extend primers hybridized to single-stranded DNA.

The uses of cDNA include the following: The cDNA synthesized by MMLV reverse transcriptase from RNA of a known sequence can be used as a template in the Polymerase Chain Reaction (PCR) for geometric amplification of a target sequence. cDNA is often used in gene cloning, or as gene probes, or in the creation of cDNA libraries.  Complementary DNA (cDNA) is DNA synthesized from a mature mRNA template in a reaction catalyzed by the enzyme reverse transcriptase. This enzyme operates on a single strand of mRNA, generating its complementary DNA based on the pairing of RNA base pairs (A, U, G and C) to their DNA complements (T, A, C and G respectively).

Often it is desirable to express eukaryotic genes in prokaryotic cells (e.g., to produce recombinant proteins in large quantities).  A simplified method of doing this involves the addition of eukaryotic DNA to a prokaryotic host, which transcribes the cDNA to mRNA and then translates it to protein.
One difference between eukaryotic and prokaryotic genes is that eukaryotic genes can contain introns, which are not coding sequences, and must be spliced out of the RNA primary transcript before it becomes mRNA and can be translated into protein. Prokaryotic genes have no introns, so their RNA is not subject to splicing

To obtain eukaryotic cDNA whose introns have been spliced: A eukaryotic cell transcribes the DNA (from genes) into RNA (pre-mRNA). The same cell processes the pre-mRNA strands by splicing out introns, and adding a poly-A tail and 5’ Methyl-Guanine cap. This mixture of mature mRNA strands is extracted from the cell. A poly-T oligonucleotide primer is hybridized onto the poly-A tail of the mature mRNA template. Reverse transcriptase requires this double-stranded segment as a primer to start its operation. The reverse transcriptase scans the mature mRNA and synthesizes a sequence of DNA that complements the mRNA template, hence the name cDNA.

USBio’s first strand cDNA is prepared by using MMLV Reverse Transcriptase.  MMLV reverse transcriptase (encoded by Moloney Murine Leukemia Virus) is an RNA-dependent DNA polymerase that synthesizes the complementary cDNA first strand from a single-stranded RNA template to which a primer has been hybridized. MMLV reverse transcriptase will also extend primers hybridized to single-stranded DNA.

The uses of cDNA include the following: The cDNA synthesized by MMLV reverse transcriptase from RNA of a known sequence can be used as a template in the Polymerase Chain Reaction (PCR) for geometric amplification of a target sequence. cDNA is often used in gene cloning, or as gene probes, or in the creation of cDNA libraries. 

Along with offering cloning products for your lab, we offer custom services. Contact tech@usbio.net for details.

Custom products
CS040-001            cDNA Library           
CS040-002            cDNA Library, Vector Preparation