고순도의 RNA 추출을 위한 Column 타입의 Takara MiniBEST 시리즈 ☐ AGPC법에 따라 total RNA를 고순도로 추출 ☐ 배양세포, 동• 식물조직 등 다양한 샘플에 적용 RNA [간단프로토콜] * 필요에 따라 homogenize TaKaRa MiniBEST Universal RNA Extraction Kit (Code 9767A) M1 : DNA Marker M2 : λ-Hind III DNA Marker gDNA M1 1 2 3 M2 TaKaRa MiniBEST Plant RNA Extraction Kit (Code 9769A) TaKaRa MiniBEST Viral RNA/DNA Extraction Kit (Code 9766A) 원하는 샘플에서 다량의 RNA를 추출하는 만점 시약 A + RNAiso Plus (Code 9109) 혈액 등의 액체샘플과 수분함유량이 많은 식물샘플에 최적 RNAiso Blood (Code 9112) ☐ Virus RNA/DNA 추출에 최적! ☐ gDNA Eraser 컬럼을 이용하여 초고순도의 RNA 추출 가능 ☐ 배양세포, 동 • 식물 조직 등 다양한 샘플의 RNA 추출에 적용 ☐ 다양한 식물세포와 조직의 RNA 추출에 최적 ☐ 잎, 종자, 과실 등 샘플의 종류에 따라 2가지 프로토콜 선택가능 Takara RNA 추출 시리즈 하나 둘 셋 액체샘플에 3배량의 RNAiso Blood를 mix* Chloroform 첨가 후 원심분리 상층 회수 후 Isopropanol 침전 고순도의 total RNA 강력추천! 1 2 3 : gDNA Eraser 컬럼 처리없이 정제한 RNA : gDNA Eraser 컬럼 처리 후 정제된 RNA : gDNA Eraser 컬럼에서 추출된 gDNA
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With PrimeSTAR GXL polymerase, products up to 30 kb could be amplified from human genomic DNA, and
products up to 40 kb were efficiently amplified from lambda DNA (Figure 2, left and middle panels). In addition,
fragments up to 13.5 kb were obtained from cDNA (Figure 2, right panel).
III. Amplification of GC-Rich Targets The performance of PrimeSTAR GXL polymerase was compared to several other commercially available high-fideli-
ty polymerases and polymerases marketed as being optimized for use with GC-rich templates.
Methods
Fragments with high GC-content were amplified from either 100 ng of human genomic DNA (lanes 1 and 2) or
10 ng of T. thermophilus HB8 genomic DNA (lanes 3 and 4). All reactions were performed according to the proto-
col recommended by each manufacturer.
Results
Under the conditions used, only PrimeSTAR GXL polymerase yielded highly specific amplification of all of the
GC-rich targets tested (Figure 3).
Conclusions
PrimeSTAR GXL polymerase outperforms other enzymes and provides highly
specific amplification in a variety of challenging PCR scenarios (i.e., long range
PCR and amplification of GC-rich sequences). With PrimeSTAR GXL polymerase,
excellent PCR results can be obtained without the need for special buffers or modifications
to the reaction conditions.
Figure 2. Amplification of long targets using PrimeSTAR GXL polymerase. Amplicons of vary-ing lengths were amplified by PCR from either human gDNA (left), lambda DNA (middle), or synthesized cDNA (right). The expected size of each product is listed beneath the gel image. In all cases, lane M contains a λ-Hind III digest molecular size marker.
M 1 2 3 4 M 1 2 3 4 M M 1 2 3 4 M 1 2 3 4 M 1 2 3 4 MM 1 2 3 4 M
PrimeSTAR GXL Company A Company B
Company B.2 (for GC-rich) Company C
Company D (for GC-rich)
Figure 3. Amplification of GC-rich targets using PrimeSTAR GXL polymerase or enzymes from other manufacturers (Companies A, B, C, or D). The Company B.2 and Company D enzymes mar-keted as optimized for GC-rich templates. Amplicons of vary-ing GC-content were amplified from either human gDNA (lane 1, APOE, 74% GC; lane 2, TBFß1, 69% GC) or T. thermophilus HB8 gDNA (lane 3, 2029 bp, 74% GC; lane 4 4988 bp, 74% GC). Lane M contains a pHY molecular size marker.