PQL-S200 (200 rxns) Store -20. using the 10-fold serial-diluted human genomic DNA and a set of human gene-specific primer

Transcription

1 CERTIFICATE OF ANALYSIS (1603-V01R02) RealHelix TM Premier q Kit [SYBR Green with low ROX] Kit contents RealHelix TM Premier q Kit [SYBR Green with low ROX] Cat. No. PQL-S200 (200 rxns) PQL-S500 (500 rxns) 2x Premier q PreMix [SL] 1 ml x 2 ea 1 ml x 5 ea Certificate Analysis 1 ea 1 ea Description The RealHelix Premier q Kit [SYBR Green with low ROX] is designed to perform a rapid real-time quantification of target DNA or 1 st -strand cdna synthesized from RNA molecule. The 2x premix contains SYBR green I dye, ROX passive dye, antibody-mediated hotstart DNA polymerase, dntps, buffers, MgCl 2, stabilizers, and enhancers. Application Quantification of target DNA sample Store -20 2x q PreMix should be stored in a dark. by real time Quality control assay data Functional analysis RealHelix Premier q Kit [SYBR Green with low ROX] was evaluated by real-time using the 10-fold serial-diluted human genomic DNA and a set of human gene-specific primer with ABI 7500 real-time system (Applied Biosystems, USA), Rotor-Gene Q instrument (QIAGEN, Germany), and Bio-Rad CFX96 (Bio-Rad, USA). Quality authorized by Youn Taek Go

2 Premier q Kit [SYBR Green with low ROX] Protocol 1. Program a real-time instrument according to the recommendations below. The excitation and emission maxima of the included SYBR Green I are at 494 nm and 521 nm, respectively. <2-step cycling protocol> If annealing temperature(at) of primers used in real-time is between 58 and 62, thermo cycling can be performed using 2-step cycling protocol as follow. Annealing & extension 60 for sec <3-step cycling protocol> If annealing temperature(at) of primers used in real-time is under 58 or above 62, thermo cycling can be performed using 3-step cycling protocol as follow. Annealing Extention AT for 30 sec 72 for 30 sec

3 Premier q Kit [SYBR Green with low ROX] 2. Add following components for a single 20 μl reaction volume.* Components DNA Template (1 ng ~ 1 μg) 2x Premier q PreMix [SL] Forward primer (10 pmoles/μl) Reverse primer (10 pmoles/μl) DEPC-treated water Volumes X μl 10 μl Adjust to final 20 μl * For multiple reactions, prepare a master mix by adding the required volumes of each above components (except the DNA template) and dispense appropriate volumes into tubes or plates. * The reaction volume for a reaction could be adjusted according to the recommendations for the instruments. 3. Gently mix and immediately centrifuge the reaction mix. 4. Perform the Real-time. Instrument compatibility 7500 Real-Time System (Applied Biosystems) CFX96 Real-Time Detection System (Bio-Rad) Rotor-Gene Q (QIAGEN) LightCycler (Roche) Mx3000P (Stratagene) Products Cat. No. Products Size PQL-S200 RealHelix Premier q Kit [SYBR Green with low ROX] 200 rxns PQL-S500 RealHelix Premier q Kit [SYBR Green with low ROX] 500 rxns

4 CERTIFICATE OF ANALYSIS (1603-V01R02) RealHelix TM Premier q Kit [SYBR Green with high ROX] Kit contents RealHelix TM Premier q Kit [SYBR Green with high ROX] Cat. No. PQH-S200 (200 rxns) PQH-S500 (500 rxns) 2x Premier q PreMix [SH] 1 ml x 2 ea 1 ml x 5 ea Certificate Analysis 1 ea 1 ea Description The RealHelix Premier q Kit [SYBR Green with high ROX] is designed to perform a rapid real-time quantification of target DNA or 1 st -strand cdna synthesized from RNA molecule. The 2x premix contains SYBR green I dye, ROX passive dye, antibody-mediated hotstart DNA polymerase, dntps, buffers, MgCl 2, stabilizers, and enhancers. Application Quantification of target DNA sample Store -20 2x q PreMix should be stored in a dark. by real-time Quality control assay data Functional analysis RealHelix Premier q Kit [SYBR Green with high ROX] was evaluated by real-time using the 10-fold serial-diluted human genomic DNA and a set of human gene-specific primer with ABI 7500 real-time system (Applied Biosystems, USA), Rotor-Gene Q instrument (QIAGEN, Germany), and Bio-Rad CFX96 (Bio-Rad, USA). Quality authorized by Youn Taek Go

5 Premier q Kit [SYBR Green with high ROX] Protocol 1. Program a real-time instrument according to the recommendations below. The excitation and emission maxima of the included SYBR Green I are at 494 nm and 521 nm, respectively. <2-step cycling protocol> If annealing temperature(at) of primers used in real-time is between 58 and 62, thermo cycling can be performed using 2-step cycling protocol as follow. Annealing & extension 60 for sec <3-step cycling protocol> If annealing temperature(at) of primers used in real-time is under 58 or above 62, thermo cycling can be performed using 3-step cycling protocol as follow. Annealing Extention AT for 30 sec 72 for 30 sec

6 Premier q Kit [SYBR Green with high ROX] 2. Add following components for a single 20 μl reaction volume.* Components DNA Template (1 ng ~ 1 μg) 2x Premier q PreMix [SH] Forward primer (10 pmoles/μl) Reverse primer (10 pmoles/μl) DEPC-treated water Volumes X μl 10 μl Adjust to final 20 μl * For multiple reactions, prepare a master mix by adding the required volumes of each above components (except the DNA template) and dispense appropriate volumes into tubes or plates. * The reaction volume for a reaction could be adjusted according to the recommendations for the instruments. 3. Gently mix and immediately centrifuge the reaction mix. 4. Perform the Real-time. Instrument compatibility 7900HT Fast Real-Time System (Applied Biosystems) 7300 Real-Time System (Applied Biosystems) 5700 Real-Time System (Applied Biosystems) StepOnePlus Real-Time System (Applied Biosystems) StepOne Real-Time System (Applied Biosystems) Products Cat. No. Products Size PQH-S200 RealHelix Premier q Kit [SYBR Green with high ROX] 200 rxns PQH-S500 RealHelix Premier q Kit [SYBR Green with high ROX] 500 rxns