Description :	dNTP Mix is an aqueous solution containing dATP, dCTP, dGTP and dTTP each 2.5mM at a final concentration of 10mM as dNTPs
Applications:	For use in PCR, cDNA synthesis, DNA sequencing and labeling procedures.
Quality Assurance: Molecular Biology Grade The dNTP Mix are certified free of nucleases (DNase, RNase), phosphateases. Each batch is validated rigorously for purity of >99% by HPLC. Each batch is tested for performance in a wide range of PCR templates with Taq and Pfu DNA polymerases. Storage and Shipping: Stable at -20℃ and shipped in dry ice ------------------------------------------------- BEAMSBIO Taq DNA Polymerase Set Taq DNA Polymerase (recombinant).  Store at –20℃ #3015S 1,000 U 1 vial of 1,000 U/vial : 5 U/µl. Supplied with 1.0ml of 10X Taq Rxn Buffer with MgCl2 and 0.5ml of 10mM dNTP mix Features ·         Generates TA cloning site of PCR products. ·         Reproducible, nuclease-free ·         PCR for General Purpose ·         RT-PCR Description Taq DNA polymerase is a heat stable DNA polymerase purified from thermophilic bacterium Thermus aquaticus.  This enzyme has an optimum temperature around 74℃ and can withstand temperatures up to 95℃.  The enzyme is unmodified and has a molecular weight of 94kDa (SDS-PAGE). Our Taq DNA polymerase is provided with x10 reaction buffer and x1 storage buffer. Our product has extremely high purity free from any nucleic acid contaminants and impurities,  which was purified through ion exchange column and affinity column chromatography. Source E. coli clone, recombinant gene from Thermus aquaticus YT1. Concentration 5 U/µl.  Successful amplification can be achieved using 1~2.5U / 50㎕ reaction, typically for PCR target < 2kb.  Further optimization, such as cycling time, reaction temperature, enzyme unit, dNTP amount, and number of cycling, will be required for PCR target greater than 2kb. Applications ·                     PCR for General Purpose ·                     TA PCR cloning ·                     Site-directed mutagenesis. Quality Control The BEAMSBIO Taq DNA Polymerase is confirmed free of endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases by strict quality tests, which shows satisfactory PCR results upto 5 kb with viral DNA templates and up to 2 kb with genomic DNA templates. Definition of Activity Unit One unit is defined as the amount of enzyme required to catalyze the incorporation of 10nmol of dNTPs into an acid-insoluble product in 30 minutes at 74℃. STANDARD UNIT ASSAY CONDITIONS  50mM Tris-HCl (pH9.0 at 25℃), 50mM NaCl, 10mM MgCl2, 200μM dATP, dCTP, dGTP, and radiolabeled dTTP, and 12.5μg activated calf thymus DNA in a 50μl reaction. Storage Buffer 50mM Tris-HCl (pH8.0), 100mM NaCl, 0.1mM EDTA, 2mM DTT, 1.0% Triton X-100 and 50% glycerol.  Store at -20℃. Inhibition and Inactivation Inactivated by phenol/chloroform extraction.  Heat denaturation is not possible. < 매뉴얼 > PCR Protocol using Taq DNA Polymerase 1. Prepare PCR tubes for the experimental samples to be amplified. 증폭할 실험시료에 적합한 PCR튜브를 준비한다. 2. Add the components in order according to following recipe : 다음 성분표를 참조하여 실험용 재료를 순서대로 넣는다.   Distilled water,                                     36~4㎕ * (balance to total vol. 50㎕)   x10 Reaction buffer,                            5㎕   10mM dNTP mix,                                 1~3㎕   Taq DNA Polymerase,                       0.5~1㎕ (2.5~5U/㎕)   DNA templates(10~200ng/㎕),          0.2~1㎕   Primer, forward(10pmol/㎕),               0.2~2㎕   Primer, backward(10pmol/㎕),           0.2~2㎕  * Primers(template와 미리 배합 사용가능)를 마지막에 넣는 것이 좋다. ---------------------------------------------------------------------------------------------------                                                             total   50㎕ 3. Mix gently and rapidly. Centrifuge down PCR tubes immediately after mixing. 가볍고 신속하게 섞고, 즉시 짧게 원심분리하여 반응물을 아래로 모은다. 4. Perform PCR using optimized cycling conditions.  Suggested cycling parameters for PCR using PCR Master Mix are : 다음 운전조건을 참조하여 PCR Master Mix 최적 PCR운전조건을 설정하여 실험을 수행한다.   - Step 1 :  initial denaturation,         94℃ for 45sec.   - Step 2 :  denaturation,                    94℃ for 45sec.                    annealing,                         Tm-5℃ for 45sec.                    extension,                         68~74℃ for 30sec~1min/kb   - Step 3 :  final extension,                 72℃ for 10min. 5. Cool down below 15℃ until harvest.   PCR Troubleshooting 1. PCR band가 전혀 없는 경우 : - 반응물 성분 배합에 착오가 없는지 확인한다.  성분누락 여부 ? - PCR 운전조건이 적합하였는지 확인한다.  PCR반응기 운전 실수 여부 ? - PCR Master Mix용도에 맞지 않는 DNA template 증폭시도 여부 ? - Primers Design 적합 여부? 2. PCR band가 뿌옇게 끌릴 때 : - Target band보다 아래쪽에서 주로 끌릴 때나 작은 non-specific band가  관찰될때는, primer design을 재검토한다.              . DNA template 불순물이 높으면 순도를 높인다.              . 길이가 길거나 증폭이 어려운 DNA template이라면 DMSO와               같은 첨가제를 적량 추가한다.  또는 Long PCR Master Mix를               선택하여 재 시도한다. ..... 이하 생략.  제품 매뉴얼 참조 .....