technical_appendix_row

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Technical Appendix Restriction Enzyme Activity in Promega 10X Buffers, Reaction Temperature and Heat Inactivation. The 10X Reaction Buffer supplied with each restriction enzyme is optimized to give 100% activity. In many cases good activity is also obtained using one of Promega’s 4-CORE® 10X Buffers. Many commonly used cloning enzymes (i.e. restriction sites are found in vector multiple cloning sites) have buffers E and H as their optimal buffer and so we have determined the activity of many of our other restriction enzymes in those buffers as well. This table may be used to select the best buffer for digestion with multiple restriction enzymes. Enzyme activity is expressed as a percent of the activity obtained with the optimal buffer for each enzyme in a one-hour digest. Enzymes with 100% activity (green) perform as well as the optimal buffer. Enzymes with 50–75% or 75–100% (yellow) will give acceptable activity in that buffer. Enzymes with <10%, 10–25%, or 25–50% (pink) activity generally do not have acceptable activity in that buffer. Also, buffers leading to star activity of the enzyme (*; pink) should be avoided. If compatible buffers cannot be identified with acceptable activity for both enzymes, each digest should be performed separately in the optimal buffer for each enzyme.

Promega Enzyme

Aat II Acc I Acc III Acc 65 I Acc B7 I Age I Alu I Alw 26 I Alw 44 I Apa I Ava I Ava II Bal I Bam H I Ban I Ban II Bbu I Bcl I Bgl I Bgl II Bsa M I Bsp 1286 I Bsr S I Bss H II Bst 98 I Bst E II Bst O I Bst X I Bst Z I Bsu 36 I Cfo I Cla I Csp I Csp 45 I Dde I Dpn I Dra I Ecl HK I Eco 47 III Eco 52 I Eco ICR I Eco R I Eco R V Fok I Hae II Hae III Hha I Hin c II Hin d III Hin f I

Cat.#

Buffer Supplied with Enzyme

Activity in A

B

R6541 R6411 R6581 R6921 R7081 R7251 R6281 R6761 R6771 R6361 R6091 R6131 R6691 R6021 R6891 R6561 R6621 R6651 R6071 R6081 R6991 R6741 R7241 R6831 R7141 R6641 R6931 R6471 R6881 R6821 R6241 R6551 R6671 R6571 R6291 R6231 R6271 R7111 R6731 R6751 R6951 R6011 R6351 R6781 R6661 R6171 R6441 R6031 R6041 R6201

J G F D E K B C C A B C G E G E A C D D D A D H D D C D D E B C K B D B B E D L B H D B B C C B E B

50–75% 50–75% <10% 10–25% 10–25% 25–50% 75–100% 10–25% <10% 100% 10–25% 50–75% 10–25% 75–100%* 25–50% 75–100% 100% 10–25% 10–25% 25–50% 10–25% 100% 10–25% 75–100% <10% 25–50% 10–25% <10% <10% <10% 75–100% 75–100% <10% 25–50% 25–50% 50–75% 75–100% <10% <10% <10% 10–25% 25–50% 10–25% 75–100% 50–75% 75–100% 50–75% 25–50% 25–50% 50–75%

10–25% 25–50% 10–25% 50–75% 50–75% 25–50% 100% 25–50% 25–50% 50–75% 100% 50–75% <10% 75–100% 25–50% 75–100% 75–100% 75–100% 25–50% 75–100% 25–50% 50–75% 25–50% 50–75% 10–25% 50–75% 25–50% 10–25% <10% 25–50% 100% 75–100% 10–25% 100% 25–50% 100% 100% <10% 25–50% <10% 100% 50–75% 25–50% 100% 100% 75–100% 75–100% 100% 100% 100%

C

D

<10% <10% 25–50% 10–25% 25–50% 25–50% 75–100% 100% 100%* <10% 25–50% 50–75% 75–100% 10–25% 100% 10–25% 100% 25–50% 50–75% <10% 50–75% 25–50% 100% 25–50% <10% <10% 75–100% 50–75% 10–25% <10% 75–100% 25–50% 75–100% <10% 100% 50–75% 75–100% 100% 75–100% 100% 50–75% 100% 25–50% 10–25% 10–25% 100% 75–100% 50–75% 10–25% 100% 50–75% 100% 100% 25–50% 25–50% 100% 10–25% 100% 50–75% 25–50% 75–100% 25–50% 100% 75–100% 25–50% 50–75% 50–75% 25–50% 50–75% 100% 75–100% 50–75% 75–100% 50–75% 75–100% 10–25% 50–75% 100% 10–25% 25–50% 75–100% <10% 50–75% 50–75% 50–75% 100% 75–100% 25–50% 50–75% 10–25% 100% 50–75% 100% 50–75% 25–50% 50–75% 75–100% 10–25% 75–100% 75–100%

www.promega.com • techserv@promega.com

E

H

MULTICORE™

10–25% <10% <10% <10% <10% 25–50% n.d. n.d. <10% 75–100% 100–125%** 100% 100% n.d. 100% n.d. n.d. 100% n.d. n.d. 10–25% n.d. n.d. 75–100% n.d. n.d. 100% 10–25% <10% 75–100% 100% 10–25% <10% n.d. n.d. 25–50% n.d. n.d. <10% 100% 50–75% 75–100% n.d. n.d. 100% n.d. n.d. 100% 10–25% 10–25% 100% 50–75% 50–75% 10–25% 25–50% 75–100% 100% n.d. n.d. <10% n.d. n.d. 25–50% n.d. n.d. 75–100% n.d. n.d. 100% n.d. 100% 75–100% n.d. n.d. 25–50% n.d. n.d. 100% n.d. n.d. <10% 100% 75–100% 10–25% 10–25% 75–100% 10–25% 100% n.d. 50–75% n.d. n.d. 100% 100% 50–75% 100% 100% 100–125%** 10–25% 100% 25–50% 50–75% n.d. n.d. 25–50% n.d. n.d. 100% n.d. n.d. 25–50% 100% n.d. 50–75% n.d. n.d. 25–50% 25–50% 50–75% <10% 25–50% n.d. 100% 75–100% 100% 100%* 25–50% 50–75% 100% n.d. n.d. 50–75% n.d. n.d. 100% n.d. n.d. 100% n.d. n.d. 75–100% 75–100% 50–75% 100% 100% 25–50% 50–75% n.d. n.d. 50–75%

Enzyme Heat Assay Inactivation Temperature + – – + + + + + + + +/– + + + – + + – + – – + – – – – – +/– – – +/– + + + +/– + + + + + + + + + – – + + + –

37°C 37°C 65°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 50°C 37°C 37°C 50°C 37°C 37°C 65°C 37°C 65°C 50°C 37°C 60°C 60°C 50°C 50°C 37°C 37°C 37°C 30°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C 37°C

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technical_appendix_row by Abdallah Abdelazeem - Issuu