Rapid detection and quantification of RNA of Ebola and Marburg viruses, Lassa virus, Crimean-Congo hemorrhagic fever virus, Rift Valley fever virus, dengue virus, and yellow fever virus by real-time reverse transcription-PCR.

Viral hemorrhagic fevers (VHFS) are acute infections with high mortality rates. Significant VHF agents are Ebola and Marburg viruses (MBGV / EBOV), Lassa Virus (LASV), Crimor Congo haemorrhagic fever virus (CCHFV), Rift Valley fever virus (RVFV), viruses dengue (DENV) and yellow fever virus (YFV). VHFs are clinically difficult to diagnose and distinguish; Fast and reliable laboratory diagnosis is required in suspect cases. We have set up six in a single step and real-time reverse-PCR transcription tests for these pathogens based on the transcriptase-platinum taq polymerase taq.

NEW-primer detection probes and / or 5′-nucleases have been designed for RVFV, DENV, YFV and CCHFV using the latest DNA database entries. PCR products have been detected in real time on a lightcycler instrument using 5′-nuclease technology (RVFV, DENV and YFV) or Sybrgreen (MBGV / EBOV, LASV and CCHFV) dye intercalation). The Sybrgreen inhibitor effect on the opposite transcription has been surmounted by an initial immobilization of the dye in the reaction capillaries. The universal cycling conditions for the detection of Sybrgreen and 5′-nuclease probe have been established. Thus, up to three tests could be carried out in parallel, facilitating rapid tests for several pathogens. All tests have been carefully optimized and validated in terms of analytical sensitivity using Vitro transcribed RNA.

The detection limits >> or = 95% determined by a probity regression analysis ranged from 1,545 to 2,835 equivalents of viral genome / ml serum (8.6 to 16 copies of RNA per test). The adequacy of the tests was illustrated by the detection and quantification of the viral RNA in serum samples of VHF patients.

Mutagenesis directed by the double-stranded DNA site by the polymerase chain reaction.

We have developed an easy procedure for the mutagenesis directed by the rapid PCR site of the DNA with double-border. The increase in the initial concentration of template and the decrease of PCR cycles to 5-10 allows us to reduce the rate of mutations of second undesirable site and considerably increase time savings. After PCR, the DPNI treatment is used to select against parental DNA molecules. The DPNI (target sequence 5′-GM6ATC) is specific to the methyl and hemimethyl DNA and is used to digest the parental DNA and select for the amplified DNA containing mutations. DNA isolated from almost all current strains Escherichia coli is the methyl dam and therefore sensitive to diNI digestion. The PFU DNA polymerase is used before the intramolecular ligation of the linear template, to remove extended bases on the 3 ‘ends of the PCR product by Taq DNA polymerase.

The recirculated vector DNA incorporating the desired mutations is transformed into E. coli. This method can be used independently of any host strain and vector. The contribution of PCR artifacts to the diversity of the ARRN genes sequence of a complex bacterioplankton sample was estimated. Taq DNA polymerase errors were found to be the dominant sequence artifact but could be forced by grouping the 99% sequence similarity group sequences. Other artifacts (chimeras and heterodeplex molecules) were significantly reduced using modified amplification protocols. Surprisingly, no falsity of sequence types have been detected in the two libraries built from amplified PCR products for different cycle numbers. Recommendations for modifying the amplification and reporting protocols of 99% diversity estimates of sequence similarity as a standard are given.

Rapid detection and quantification of RNA of Ebola and Marburg viruses, Lassa virus, Crimean-Congo hemorrhagic fever virus, Rift Valley fever virus, dengue virus, and yellow fever virus by real-time reverse transcription-PCR.
Rapid detection and quantification of RNA of Ebola and Marburg viruses, Lassa virus, Crimean-Congo hemorrhagic fever virus, Rift Valley fever virus, dengue virus, and yellow fever virus by real-time reverse transcription-PCR.

Enzymatic assembly of overlapping DNA fragments.

Three methods of assembling several DNA molecules overlapping are described. Each method shares the same basic approach: (i) an exonuclease eliminates the nucleotides of the ends of the dual-fired DNA molecules (DS), exposing specific advertising DNA (SS) overhangs; (ii) SSDNA gaps of the joined molecules are filled by DNA polymerase and pseudonyms are sealed covalently by DNA ligase. The first method uses the 3′-exonuclease of T4 DNA polymerase (T4 POL), Taq DNA polymerase (Taq Pol) and the Taq Taq Ligase (Taq Lig) in a two-step thermocyclic reaction. The second method uses 3′-exonuclease III (EXOIII), Taq Pol and Taq Antibody Lag in a thermocyclic reaction in one step.

The third method uses the 5′-T5 exonuclease, the polymerase of the DNA phusion® and the TAQ LIG in an isothermal reaction in one step and can be used to assemble both SSDNA and DSDNA. These assembly methods can be used to transparently construct synthetic and natural genes, genetic pathways and integer genomes and could be very useful for molecular engineering tools. The nucleotide sequences in three hypervarial regions of the Human Immunodeficiency Virus (HIV-1) will envisage by provirus sequencing present in peripheral blood mononuclear cells of individuals infected with HIV. Simple molecules of target sequences have been isolated by limiting dilution and amplified in two steps by reaction of the polymerase chain, using nested primers.

RNA polymerase II RPB11 Antibody

GWB-81535B 0.05 ml Ask for price

TAF9 RNA Polymerase II (Recombinant)

20-abx073191
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  • 1 mg
  • 20 ug
  • 5 ug

TAF10 RNA Polymerase II (Recombinant)

20-abx073836
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  • 10 ug
  • 1 mg
  • 2 µg

RNA Polymerase II(CTD4H8), 0.2mg/mL

BNUB2929-100 1uL
EUR 225
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

RNA Polymerase II(CTD4H8), 0.2mg/mL

BNUB2929-500 1uL
EUR 485
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

RNA Polymerase II(CTD4H8), 1mg/mL

BNUM2929-50 1uL
EUR 396
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Recombinant Human TAF9 RNA Polymerase II

7-03625 5µg Ask for price

Recombinant Human TAF9 RNA Polymerase II

7-03626 20µg Ask for price

Recombinant Human TAF9 RNA Polymerase II

7-03627 1mg Ask for price

TAF10 RNA Polymerase II Human Recombinant

rAP-1598 Inquiry Ask for price

RNA Polymerase II Monoclonal Antibody [CTD4H8]

A-2032
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  • 100 µg
  • 50 ug
  • 100 ug

RNA Polymerase II Subunit B1 Mouse mAb

E2222646 100ul
EUR 255
Description: Biotin-Conjugated, FITC-Conjugated , AF350 Conjugated , AF405M-Conjugated ,AF488-Conjugated, AF514-Conjugated ,AF532-Conjugated, AF555-Conjugated ,AF568-Conjugated , HRP-Conjugated, AF405S-Conjugated, AF405L-Conjugated , AF546-Conjugated, AF594-Conjugated , AF610-Conjugated, AF635-Conjugated , AF647-Conjugated , AF680-Conjugated , AF700-Conjugated , AF750-Conjugated , AF790-Conjugated , APC-Conjugated , PE-Conjugated , Cy3-Conjugated , Cy5-Conjugated , Cy5.5-Conjugated , Cy7-Conjugated Antibody

RNA Polymerase II Subunit B1 Rabbit mAb

E2R26223 100ul
EUR 255
Description: Available in various conjugation types.

RNA Polymerase II Subunit B3 Rabbit mAb

E2R389083 100ul
EUR 275
Description: Biotin-Conjugated, FITC-Conjugated , AF350 Conjugated , AF405M-Conjugated ,AF488-Conjugated, AF514-Conjugated ,AF532-Conjugated, AF555-Conjugated ,AF568-Conjugated , HRP-Conjugated, AF405S-Conjugated, AF405L-Conjugated , AF546-Conjugated, AF594-Conjugated , AF610-Conjugated, AF635-Conjugated , AF647-Conjugated , AF680-Conjugated , AF700-Conjugated , AF750-Conjugated , AF790-Conjugated , APC-Conjugated , PE-Conjugated , Cy3-Conjugated , Cy5-Conjugated , Cy5.5-Conjugated , Cy7-Conjugated Antibody

RNA Polymerase II Subunit J (POLR2J) Antibody

20-abx325624
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  • 100 ug
  • 50 ug

RNA Polymerase II Subunit C (POLR2C) Antibody

20-abx318103
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  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

RNA Polymerase II Subunit D (POLR2D) Antibody

20-abx304886
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  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

RNA Polymerase II Subunit E (POLR2E) Antibody

20-abx328881
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  • 100 ug
  • 50 ug

RNA Polymerase II Subunit J (POLR2J) Antibody

20-abx306636
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  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

RNA Polymerase II Subunit M (POLR2M) Antibody

20-abx333780
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  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

RNA Polymerase II Subunit I (POLR2I) Antibody

20-abx301254
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  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

RNA Polymerase II Subunit A (POLR2A) Antibody

20-abx330038
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  • 100 ug
  • 50 ug

RNA Polymerase II Subunit C (POLR2C) Antibody

abx034956-400ul 400 ul
EUR 627.6

RNA Polymerase II Subunit C (POLR2C) Antibody

abx034956-80l 80 µl
EUR 343.2

RNA Polymerase II Subunit C (POLR2C) Antibody

abx036033-100ug 100 ug
EUR 469.2

RNA Polymerase II Subunit H (POLR2H) Antibody

abx036051-100ug 100 ug
EUR 469.2

RNA Polymerase II Subunit J (POLR2J) Antibody

abx036622-100ug 100 ug
EUR 469.2

RNA Polymerase II Subunit F (POLR2F) Antibody

20-abx123154
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  • 100 ul
  • 200 ul
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RNA Polymerase II Subunit L (POLR2L) Antibody

20-abx123460
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  • 100 ul
  • 200 ul
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RNA Polymerase II Subunit M (POLR2M) Antibody

20-abx126390
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  • 100 ul
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RNA Polymerase II Subunit H (POLR2H) Antibody

20-abx127012
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  • 100 ul
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RNA Polymerase II Subunit C (POLR2C) Antibody

abx236621-100ug 100 ug
EUR 577.2

RNA Polymerase II Subunit D (POLR2D) Antibody

abx236622-100ug 100 ug
EUR 610.8

RNA Polymerase II Subunit F (POLR2F) Antibody

abx236623-100ug 100 ug
EUR 577.2

RNA Polymerase II Subunit H (POLR2H) Antibody

abx236624-100ug 100 ug
EUR 577.2

RNA Polymerase II Subunit I (POLR2I) Antibody

abx236625-100ug 100 ug
EUR 610.8

RNA Polymerase II Subunit J (POLR2J) Antibody

abx236626-100ug 100 ug
EUR 577.2

RNA Polymerase II Subunit L (POLR2L) Antibody

abx236628-100ug 100 ug
EUR 610.8

RNA Polymerase II Subunit L (POLR2L) Antibody

abx218226-100ug 100 ug
EUR 526.8

RNA Polymerase II Subunit C (POLR2C) Antibody

20-abx142152
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  • 100 ul
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RNA Polymerase II Subunit D (POLR2D) Antibody

20-abx142153
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  • 100 ul
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  • 50 ul

RNA Polymerase II Subunit D (POLR2D) Antibody

abx145396-100ug 100 ug
EUR 469.2

RNA Polymerase II Subunit B (POLR2B) Antibody

20-abx114601
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  • 150 ul
  • 50 ul

RNA Polymerase II Subunit C (POLR2C) Antibody

20-abx114602
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  • 150 ul
  • 50 ul

RNA Polymerase II Subunit I (POLR2I) Antibody

20-abx114607
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  • 150 ul
  • 50 ul

RNA Polymerase II Subunit L (POLR2L) Antibody

20-abx114608
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  • 150 ul
  • 50 ul

RNA Polymerase II Subunit B (POLR2B) Antibody

20-abx004544
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  • 100 ul
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RNA Polymerase II Subunit H (POLR2H) Antibody

20-abx005271
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  • 100 ul
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RNA Polymerase II Subunit I (POLR2I) Antibody

20-abx008848
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  • 100 ul
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  • 30 ul

RNA Polymerase II Subunit H (POLR2H) Antibody

20-abx008849
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  • 100 ul
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  • 30 ul

RNA Polymerase II Subunit C (POLR2C) Antibody

20-abx001481
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  • 100 ul
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  • 20 ul
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RNA Polymerase II Subunit F (POLR2F) Antibody

20-abx001507
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  • 100 ul
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RNA Polymerase II Subunit J (POLR2J) Antibody

20-abx001523
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  • 100 ul
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RNA Polymerase II Subunit D (POLR2D) Antibody

20-abx001535
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  • 100 ul
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  • 20 ul
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RNA Polymerase II Subunit I (POLR2I) Antibody

20-abx001539
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  • 100 ul
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  • 20 ul
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RNA Polymerase II Subunit L (POLR2L) Antibody

20-abx001935
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  • 100 ul
  • 200 ul
  • 20 ul
  • 50 ul

RNA Polymerase II Subunit H (POLR2H) Antibody

abx031477-400ul 400 ul
EUR 627.6

RNA Polymerase II Subunit H (POLR2H) Antibody

abx031477-80l 80 µl
EUR 343.2

RNA Polymerase II Subunit J (POLR2J) Antibody

abx027917-400ul 400 ul
EUR 627.6

RNA Polymerase II Subunit J (POLR2J) Antibody

abx027917-80l 80 µl
EUR 343.2

RNA Polymerase II Subunit B (POLR2B) Antibody

abx028262-400ul 400 ul
EUR 627.6

RNA Polymerase II Subunit B (POLR2B) Antibody

abx028262-80l 80 µl
EUR 343.2

RNA Polymerase II Subunit D (POLR2D) Antibody

abx029360-400ul 400 ul
EUR 627.6

RNA Polymerase II Subunit D (POLR2D) Antibody

abx029360-80l 80 µl
EUR 343.2

RNA Polymerase II Subunit H (POLR2H) Antibody

abx005271-100l 100 µl
EUR 275

RNA Polymerase II Subunit H (POLR2H) Antibody

abx005271-200l 200 µl
EUR 400

RNA Polymerase II Subunit H (POLR2H) Antibody

abx005271-50l 50 µl
EUR 175

RNA Polymerase II Subunit C (POLR2C) Antibody

abx001481-100l 100 µl
EUR 400

RNA Polymerase II Subunit C (POLR2C) Antibody

abx001481-20l 20 µl
EUR 175

RNA Polymerase II Subunit C (POLR2C) Antibody

abx001481-50l 50 µl
EUR 275

RNA Polymerase II Subunit F (POLR2F) Antibody

abx001507-100l 100 µl
EUR 387.5

RNA Polymerase II Subunit F (POLR2F) Antibody

abx001507-20l 20 µl
EUR 175

RNA Polymerase II Subunit F (POLR2F) Antibody

abx001507-50l 50 µl
EUR 275

RNA Polymerase II Subunit J (POLR2J) Antibody

abx001523-100l 100 µl
EUR 400

RNA Polymerase II Subunit J (POLR2J) Antibody

abx001523-20l 20 µl
EUR 175

RNA Polymerase II Subunit J (POLR2J) Antibody

abx001523-50l 50 µl
EUR 275

RNA Polymerase II Subunit D (POLR2D) Antibody

abx001535-100l 100 µl
EUR 400

RNA Polymerase II Subunit D (POLR2D) Antibody

abx001535-20l 20 µl
EUR 175

RNA Polymerase II Subunit D (POLR2D) Antibody

abx001535-50l 50 µl
EUR 275

RNA Polymerase II Subunit I (POLR2I) Antibody

abx001539-100l 100 µl
EUR 400

RNA Polymerase II Subunit I (POLR2I) Antibody

abx001539-20l 20 µl
EUR 175

RNA Polymerase II Subunit I (POLR2I) Antibody

abx001539-50l 50 µl
EUR 275

RNA Polymerase II Subunit J (POLR2J) Antibody

abx306636-100g 100 µg
EUR 362.5

RNA Polymerase II Subunit J (POLR2J) Antibody

abx306636-20g 20 µg
EUR 162.5

RNA Polymerase II Subunit J (POLR2J) Antibody

abx306636-50g 50 µg
EUR 250

RNA Polymerase II Subunit D (POLR2D) Antibody

abx304886-100g 100 µg
EUR 362.5

RNA Polymerase II Subunit D (POLR2D) Antibody

abx304886-20g 20 µg
EUR 162.5

RNA Polymerase II Subunit D (POLR2D) Antibody

abx304886-50g 50 µg
EUR 250

RNA Polymerase II Subunit C (POLR2C) Antibody

abx236621-100g 100 µg
EUR 350

RNA Polymerase II Subunit D (POLR2D) Antibody

abx236622-100g 100 µg
EUR 350

RNA Polymerase II Subunit F (POLR2F) Antibody

abx236623-100g 100 µg
EUR 350

RNA Polymerase II Subunit H (POLR2H) Antibody

abx236624-100g 100 µg
EUR 350

RNA Polymerase II Subunit I (POLR2I) Antibody

abx236625-100g 100 µg
EUR 350

RNA Polymerase II Subunit J (POLR2J) Antibody

abx236626-100g 100 µg
EUR 350

RNA Polymerase II Subunit I (POLR2I) Antibody

abx301254-100g 100 µg
EUR 362.5

RNA Polymerase II Subunit I (POLR2I) Antibody

abx301254-20g 20 µg
EUR 162.5

RNA Polymerase II Subunit I (POLR2I) Antibody

abx301254-50g 50 µg
EUR 250

RNA Polymerase II Subunit C (POLR2C) Antibody

abx114602-100l 100 µl
EUR 612.5

RNA Polymerase II Subunit I (POLR2I) Antibody

abx114607-100l 100 µl
EUR 612.5

RNA Polymerase II Subunit D (POLR2D) Antibody

abx145396-1096tests 10 × 96 tests Ask for price

RNA Polymerase II Subunit D (POLR2D) Antibody

abx145396-596tests 5 × 96 tests Ask for price

RNA Polymerase II Subunit D (POLR2D) Antibody

abx145396-96tests 96 tests
EUR 337.5

RNA Polymerase II Subunit C (POLR2C) Antibody

abx034956-100g 100 µg
EUR 281.25

RNA Polymerase II Subunit C (POLR2C) Antibody

abx036033-100g 100 µg
EUR 337.5

RNA Polymerase II Subunit H (POLR2H) Antibody

abx036051-100g 100 µg
EUR 337.5

RNA Polymerase II Subunit D (POLR2D) Antibody

abx029360-400l 400 µl
EUR 518.75

RNA Polymerase II Subunit J (POLR2J) Antibody

abx036622-100g 100 µg
EUR 337.5

RNA Polymerase II Subunit H (POLR2H) Antibody

abx031477-400l 400 µl
EUR 518.75

RNA Polymerase II Subunit F (POLR2F) Antibody

abx123154-100l 100 µl
EUR 175

RNA Polymerase II Subunit F (POLR2F) Antibody

abx123154-1ml 1 ml
EUR 400

RNA Polymerase II Subunit F (POLR2F) Antibody

abx123154-200l 200 µl
EUR 275

RNA Polymerase II Subunit J (POLR2J) Antibody

abx027917-400l 400 µl
EUR 518.75

RNA Polymerase II Subunit J (POLR2J) Antibody

abx325624-100g 100 µg
EUR 250

RNA Polymerase II Subunit J (POLR2J) Antibody

abx325624-50g 50 µg
EUR 187.5

RNA Polymerase II Subunit A (POLR2A) Antibody

abx330038-100g 100 µg Ask for price

RNA Polymerase II Subunit A (POLR2A) Antibody

abx330038-20g 20 µg
EUR 187.5

RNA Polymerase II Subunit A (POLR2A) Antibody

abx330038-50g 50 µg
EUR 250

RNA Polymerase II Subunit M (POLR2M) Antibody

abx333780-100g 100 µg
EUR 362.5

RNA Polymerase II Subunit M (POLR2M) Antibody

abx333780-20g 20 µg
EUR 162.5

RNA Polymerase II Subunit M (POLR2M) Antibody

abx333780-50g 50 µg
EUR 250

RNA Polymerase II Subunit E (POLR2E) Antibody

abx328881-100l 100 µl
EUR 187.5

RNA Polymerase II Subunit C (POLR2C) Antibody

abx318103-100g 100 µg
EUR 362.5

RNA Polymerase II Subunit C (POLR2C) Antibody

abx318103-20g 20 µg
EUR 162.5

RNA Polymerase II Subunit C (POLR2C) Antibody

abx318103-50g 50 µg
EUR 250

T7 RNA Polymerase

E041 100 μl (5000 U)
EUR 65
Description: T7 RNA Polymerase is a DNA dependent RNA polymerase that catalyzes the synthesis of RNA in the 5´→ 3´ direction only in the presence of its cognate T7 phage promoter sequence. T7 RNA Polymerase has high specificity for the T7 phage promoter and will not recognize SP6 or T3 RNA Polymerase promoter sequences.

T7 RNA Polymerase

K1083-100000 100000U
EUR 512
Description: DNA-Dependent RNA Polymerase Specific for T7 Promoter Sequences

T7 RNA Polymerase

K1083-25000 25000U
EUR 160
Description: DNA-Dependent RNA Polymerase Specific for T7 Promoter Sequences

T7 RNA Polymerase

K1083-5000 5000U
EUR 40
Description: DNA-Dependent RNA Polymerase Specific for T7 Promoter Sequences

T7 RNA Polymerase

PCR-603L 5 x 4000 units
EUR 251.6

T7 RNA Polymerase

PCR-603S 4000units
EUR 63

anti-RNA Polymerase II p14.5

YF-PA24419 50 ul
EUR 400.8
Description: Mouse polyclonal to RNA Polymerase II p14.5

RNA Polymerase II Subunit J2 (POLR2J2) Antibody

abx236627-100ug 100 ug
EUR 661.2

The product was directly sequenced to avoid errors introduced by Taq polymerase during the amplification process. There was an extended variation between the sequences of the same individual as between the sequences of different individuals. Interpanding variability was significantly lower in people infected with a common source. A high proportion of amino acid substitutions in hypervariable regions alter the number and positions of potential glycosylation sites n-linked.