High Efficiency GC5 Competent cells

COMMENTS: 
The transformation efficiency of GC5ä Standard Aliquots is 1 x 109 cfu/µg pUC 19 plasmid DNA. pUC 19 Control DNA is provided.

APPLICATION:

GC5 chemically competent cells are comparable to the popular DH5a strain and carry recA1 and endA1 mutations that aid in plasmid stability and improved quality of prepared plasmid DNA. The GC5 strain carries the beta-galactosidase a-complementation factor for blue/white screening and are T1 bacteriophage resistant. GC5 Competent Cells are offered as a high efficiency grade for subcloning and generation of cDNA libraries.

UptiTherm EC DNA Polymerase

The advantages of a native polymerase with the reproducibility of a recombinant enzyme.

FEATURES:

• Thermostable DNA polymerase from Thermus thermophilus. Expressed in recombinant form in E. coli

• Purified in order to remove all traces of genomic DNA

• The purified enzyme possesses 5’- 3’ polymerase activity, as well as weak 5’-3’ exonuclease activity. No 3’-5’ activity is detected (it is not a proof-reading enzyme). The enzyme does not present significant reverse transcriptase activity

• Lower error rates than Taq (Taq error rate 1/6 Kbp, Tth error rate 1/10 Kbp)

• Half life at 94 ºC is 40 min. Polymerase activity remains unaltered after more than 40 cycles of amplification

• Radioactively labelled dNTPs, as well as biotin, fluorescein and digoxigenin-labelled dNTPs (including dUTP) can be used as substrates

• Reaction products can be directly used in T/A cloning

APPLICATIONS:

• Routine amplifications involving sequences homologous to E. coli.

• Amplifications involving a high number of cycles (up to 55-60)

• Amplifications up to 5 Kbp

• Amplifications involving genomic DNA

• Amplifications with limiting amounts of template

• Plasmid amplifications (including direct colony screening)

• Multiplex amplifications

• Tested in RAPDs and DNA chips

Figure 1.

Genomic DNA from different E. coli strains (lanes 1 to 4) was amplified using a set of random decamers, at an annealing temperature of 35 ºC. Lanes 5 and 6 are negative controls. M: 100 bp molecular ladder (Cat. No. N15101).

UptiPfu DNA Polymerase,  1 U/ µl

Catalog #

Product

AK5100

100 units + Standard buffer

AK5101

250 units + Standard buffer

AK5102

500 units + Standard buffer

AK5103

100 units + MgCl₂ free buffer>

AK5104

250 units + MgCl₂ free buffer

AK5105

500 units + MgCl₂ free buffer

Components of the standard kit:

• 15 ml MELT solution

• 1 ml BIND solution

• 3 ml 40 x WASH solution

(for 120 ml WASH solution mix with 60 ml ethanol and 57 ml H₂O)

The purified DNA can be used for digestion with restriction enzymes, for ligation, transformation and labeling with Klenow enzyme. DNA fragments from 20 bp up to 10 kb length can be successfully purified with this kit.