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商品編號


RT80125K



品牌


Lucigen



公司


Lucigen



公司分類(lèi)


Reverse Transcriptase Enzymes



Size

25,000 Units






商品信息

Optimized reverse transcriptase and buffer system for the production of full-length
CDN
A.


Synthesize full-length
CDN
A (>15 kb)

Amplify first-strand
CDN
A from picogram amounts of total RNA

Optimize the RT reaction to your specific needs by using the first strand
CDN
A synthesis primers, dNTPs and RNase inhibitor of your choice (not included).





The MMLV HP RT demonstrates significantly greater reverse transcriptase activity than other commercially available MMLV RT enzymes. Typically, just 100 units of MMLV HP RT are required for full-length
CDN
A synthesis compared to 200 units of competitive MMLV RT enzymes. MMLV HP RT includes a 10X Reaction Buffer, optimized for synthesis of full-length
CDN
A from long RNA templates, and DTT. The enzyme, buffer and DTT are the same components used in the MMLV Reverse Transcriptase 1st-Strand
CDN
A Synthesis Kit. By providing these three components individually, you have the flexibility to choose dNTPs, RNase Inhibitors, etc. and optimize the RT reaction for your specific needs.


Figure 1. MMLV HP RT produces full-length
CDN
A from mRNA longer than 15 kb.
?Total RNA isolated from HeLa cells was reverse transcribed and the
CDN
A was amplified by PCR. Detection of the 1.3-kb PCR amplicon from near the 5? end of the mRNA demonstrates full-length reverse transcription of HERC1 mRNA (A). Agarose gel analysis of the PCR products shows the 1.3-kb amplicon from the 5? end of the mRNA (B). Lane M, 100 bp DNA ladder; lane 1, no-RT control reaction; lane 2, PCR product from
CDN
A synthesized by Epicentre's MMLV HP RT.





Target Transcript (Size)

3?/5? Ratios



Epicentre MMLV High Performance Reverse Transcriptase

Competitor I (RNase H-Mutant of MMLV RT)

Competitor P (MMLV RT)



ACTB (1,792 b)

0.9

1.7

1.2



GUSB (2,162 b)

1.0

6.1

2.5



TFRC (5,010 b)

5.5

12.1

11.3





Table 1. 3?/5? ratio analysis of
CDN
A produced by different reverse transcriptase enzymes.
?Total cellular RNA from HeLa cells was converted to
CDN
A using the three reverse transcriptase enzymes indicated in the table. A 3?/5? ratio equal to 1.0 means that equal amounts of PCR products are obtained from both the 3? and 5? end of the
CDN
A and therefore is a good indication that the reverse transcriptase has produced a full-length
CDN
A copy of the mRNA.


2A.


2B.

Epicentre MMLV 1st-Strand
CDN
A Synthesis Kit


Company I RNase H- minus MMLV RT


Company P MMLV RT


Figure 2.
CDN
A produced by Epicentre's MMLV High Performance Reverse Transcriptase yields a significantly improved 3?/5? ratio than competitive reverse transcriptases.
?The approximately 2160-base HeLa β-glucuronidase mRNA (GUSB) was reverse transcribed into
CDN
A using Epicentre's MMLV High Perfomance Reverse Transcriptase and two competitive reverse transcriptase enzymes. PCR primer pairs to the 3?-end and 5?-end of GUSB
CDN
A were synthesized and qPCR (SYBR??Green I dye detection) was performed using each primer pair and the GUSB
CDN
As as templates. The 3?/5? ratio was calculated for each as described in the text.
(A).
PCR amplicons from the 3? end and 5? end of the GUSB
CDN
A.
(B)
. qPCR quantification graphs for detecting the 3? amplicon and 5? amplicon of GUSB
CDN
A produced by Epicentre's MMLV High Performance Reverse Transcriptase Kit and two other commercially available reverse transcriptase enzymes.

ORDER INFORMATION
MMLV High Performance Reverse Transcriptase, 10X Reaction Buffer, DTT.