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PCR Prep PCR Workstation

Item #: ACRPCRHOOD4 
Bullseye dNTP Set (100mM)


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RT 2X Master Mix

  • Up to 9kb cDNA synthesis
  • Ensures sample to sample consistency
  • Large RNA sample volume capacity
  • Ready to use

Size: 100 rxns

RT 2X Master Mix is a proprietary, ready-to-use master mix for first-strand cDNA synthesis in a 2X concentration. This optimized reaction mix contains ribonuclease inhibitor, dNTPs, and a balanced concentration for oligo(dT) and random primers. The ribonuclease inhibitor effectively protects RNA template from degradation. The oligo(dT) anneals selectively to the poly(A) tail of mRNAs and the random primers do not require the presence of poly(A) and they are utilized for the transcription of mRNA 5-end regions. The resultant cDNA can be directly used as template in different PCR experiments.

Kit Components

EasyScript R Tase (200U / uL)100 L
2X Reaction Mix1200 uL
Nuclease-Free H2O2 x 1 mL

Storage

  • Store at -20°C in a frost-free freezer.

Item #DescriptionQuantityRxn
BERTCDNA-25RT 2X Master Mix  250 uL25 rxns
BERTCDNA-100RT 2X Master Mix 1 mL100 rxns


Application

  • cDNA synthesis
  • Construction of cDNA libraries
  • Generation of probes for hybridization 

Protocol

  1. Thaw RNA templates and all reagents on ice. Mix each solution by vortexing.
  2. Assemble the following components in a tube on ice, and mix well:

ComponentsVolumeFinal Conc.
Total RNA, orVariable1 ng - 2 ug/rxn
mRNAVariable1 pg - 2 ng/rxn
2X Reaction Mix10 uL1X
H2OUp to 19 uL-

  1. Heat the mixture at 65°C for 5 mins and incubate on ice for at least 1 min.
  2. Collect all components by a brief centrifugation and add 1 µl of the EasyScript RTase to the tube.
  3. Mix well and collect all the components by a brief centrifugation.
  4. Incubate the tube at room temperature for 10 min for annealing.
  5. Perform cDNA synthesis by incubating the tube for 50 min at 42°C.
  6. Stop the reaction by heating it at 85°C for 5 min.
  7. Chill on ice. The newly synthesized first-strand cDNA is ready for immediate downstream applications. 

PNCDNART

Pkg
Master Mix
PR1MA Enzyme Container
Item#:
ASPRIMART
 
One-step RT-PCR Kit
Item#:
ASRTPCR
 
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PCR Prep PCR Workstation
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