pcr cloning protocols - harry w janes, bing-yuan chen

... Func- tional Proteomics, edited by Christoph Kannicht, 2002 193. RT -PCR Protocols, edited by Joseph O’Connell, 2002 192. PCR Cloning Protocols, 2nd ed., edited by Bing-Yuan Chen and Harry W. Janes, ... Pretreatment of each PCR reaction with uracil-N glycosylase (UNG), which catalyzes the removal of uracil from single- and double- PCR: Basic Principles 5 stranded DNA, will...

Ngày tải lên: 08/04/2014, 12:52

... 6.20,6.21,9.1 6-9 ,19,9.3 4-9 .57, and B.23,B.24. 11. Chou, Q., Russell, M., Birch, D. E., Raymond, J., and Bloch, W. (1992) Preven- tion of pre -PCR mis-priming and primer dimerization improves low-copy-num- ... sites. 2. Two positive control primer sets for XL PCR with human genomic DNA are listed below (5 -3 ’). These primers were designed for use with a 6 7-6 8’C annealing...

Ngày tải lên: 11/04/2014, 10:01

... products with another 30 cycles of PCR and isolate the product for sequencing again as described m the protocol. We have not tested whether a PCR- based cycle sequencing method would work well with ... mobility were seen whether the 5 - or 3’ -PCR primer was used as a sequencing primer, suggesting that some factor other than secondary structure or reannealing of the PCR product wa...

Ngày tải lên: 11/04/2014, 10:02

... organism. However, the safest approach is to design primers that reflect all possible Cloning genes by PCR 249 (A) (B) Coding region N-term Pep-1 Pep-2 Coding region N-term Pep-2 Pep-1 Figure 10.11 Primer ... primer for first-strand cDNA synthesis and homopolymer-tailing, often by dCTP, of the 3′-end of these cDNA strands. PCR with oligo-dG and oligo-dT primers was then performed. This ap...

Ngày tải lên: 25/10/2013, 22:20

... product GCCTACTACCATC CGGATGATGGTAG GATGGTAGTAGGC CGGATGATGGTAG CGATGGTAGTAGG GCTACCATCATCC GCTACCATCATCC CCTACTACCATCG CGATGGTAGTAGGC GCTACCATCATCCG GCCTACTACCATCG CGGATGATGGTAGC PCR amplify vector using primers with sequence-specific tails Digest 3'-ends with T4 DNA polymerase and dCTP Anneal PCR product from (A) with vector from (B) PCR- amplified product PCR- amplified ... succ...

Ngày tải lên: 25/10/2013, 22:20

... follows.  Consider now the mapping w =e z .By(5), we have w =e x (cos y +isin y), where x, y ∈ R.It follows that |w| =e x and arg w = y +2πk, where k ∈ Z. Usually we make the choice arg w = y, with ... Arg (w) , where Arg (w) denotes the principal argument of w. Itfollows that x = log |w| and y = Arg (w) . Hence (10) Log (w) =log |w| +iArg (w) . In many practical situations, we usu...

Ngày tải lên: 31/03/2014, 15:17

... for the wild-type enzyme. The catalytic ef®ciency (k cat /K m )for D -xylose for M- 1026 was 6% higher than that of the wild-type, while for the other mutants it was lower. As for the wild-type ... Three cold-adapted mutants were isolated with the following amino-acid substitutions: E372G, V379A (M-1021), E372G, F163L (M-1024) and E372G (M-1026). The wild- type and mutated xylA genes w ere...

Ngày tải lên: 31/03/2014, 15:20

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Ngày tải lên: 31/03/2014, 15:56

... http://research. nwfsc.noaa.gov /protocols/ methods/ 2. http://www.stratagene.com/ 3. http://www.promega.com/tbs/ 4. http://www.highveld.com /protocols. html 5. http://www.dynal.no/ 6. Sambrook, J., ... unidirectional workow (4). This avoids back ow of trafc and, along with restricted access, will reduce the risk of contamination and inaccurate results. The way in which the workow...

Ngày tải lên: 08/04/2014, 12:52