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BioForum > Methods Discussion > Molecular Cloning
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  1. Pinned: What is the biggest insert for a viral vector? (14 replies)
  2. Pinned: Creating empty vector by self-ligation (31 replies)
  3. Pinned: Time needed to determine G418 concentration for selection (5 replies)
  4. Pinned: Smear in my restriction digest (11 replies)
  5. Pinned: How to transfect two plasmids in a cell (10 replies)
  6. Pinned: ds-oligo cloning/ligation issues (8 replies)
  7. Pinned: Sequential restriction digestion of plasmid (51 replies)
  8. Pinned: Problems with Stratagene Site Directed Mutagenesis (21 replies)
  9. Pinned: Help with 3-way ligation (11 replies)
  10. Pinned: Old threads on Molecular Cloning are achived here (0 replies)
  11. pUCP18 LacZ alfa complementation (0 replies)
  12. keeping restriction digested fragments at -20? (0 replies)
  13. Need help with pkD46, Pkd13, pCP20 (0 replies)
  14. re-digestion (1 reply)
  15. Freezing PCR products prior to ligation (2 replies)
  16. Cloning woes - Running out of ideas! (7 replies)
  17. Has anyone worked with pCHF3 (0 replies)
  18. Have you ever use pDM4 plasmid (0 replies)
  19. Cloning using BamH1 and HindIII (1 reply)
  20. Gel purification of vector DNA - low yield (26 replies)
  21. frozen competent cells (2 replies)
  22. plasmid extraction kit wash buffer solution (3 replies)
  23. minimum amounts of insert and vector for ligation (4 replies)
  24. Screening colonies after transformation (2 replies)
  25. "competentcy" of my method (0 replies)
  26. Fission Yeast complementaion (0 replies)
  27. Vector frameshift problem (3 replies)
  28. Huge difference in Tm of my For and Rev primers = no PCR product?!? (11 replies)
  29. z competent cells (2 replies)
  30. insert release not as intense as the plasmid (3 replies)
  31. Cell culture staining (0 replies)
  32. partial DNA digestion formula (1 reply)
  33. molecular "terminology" (1 reply)
  34. Sequence management software (for Mac) (4 replies)
  35. NdeI digestion (3 replies)
  36. Cloning IRES (0 replies)
  37. Multiple bands after amping insert (1 reply)
  38. Good tool for sequence validation? (2 replies)
  39. problem in plasmid isolation (5 replies)
  40. pCDH Lentivector has 500 bp insert (4 replies)
  41. screening of positive clones (3 replies)
  42. Insertion Mutation in Insert Sequence (2 replies)
  43. Small fragment (90bp) into an AAV backbone (4.7kb) ligation problem (1 reply)
  44. Silencing of the first cistron (1 reply)
  45. To clean or not to clean.....? (3 replies)
  46. Neon Transfection System (2 replies)
  47. why they use isopropanal for precipitate DNA in midiprep (5 replies)
  48. Stable Mammalian Expression with Retrovirus (2 replies)
  49. help (5 replies)
  50. Subcloning promoter region from one vector to another (0 replies)
  51. Correct orientation of the insert (7 replies)
  52. problems with qiagen gel extraction kit (7 replies)
  53. Molecular Cloning (1 reply)
  54. restriction enzyme flanking sequence preferance (2 replies)
  55. 5' RACE for very GC-rich gene (2 replies)
  56. How to amplify unknown cDNA sequences from a RIP for sequencing (1 reply)
  57. Phosphorylated primers (5 replies)
  58. Cloning advice needed (4 replies)
  59. PIR 1 Competent Cells (0 replies)
  60. help (1 reply)
  61. Colony Screeing without using PCR (8 replies)
  62. pcr amolification with long pcr enzyme mix for cloning (1 reply)
  63. blue white screening (3 replies)
  64. Odd point mutations (23 replies)
  65. phosphorylate dprimers for pfx do not work (3 replies)
  66. problem with yeast transformation (0 replies)
  67. ta cloning of pfu amplified fragment (6 replies)
  68. how to express the gene in opposite transcriptional direction (3 replies)
  69. Water for electrocompetent cells (2 replies)
  70. Luciferase GL4 siRNA (5 replies)
  71. Bacterial growth in agar plate with ampicillin (4 replies)
  72. problem with 293T cell co-transfection (5 replies)
  73. Spike Pfu's for quikchange mutagenesis? (0 replies)
  74. AAARGHHH, cloning driving me insane, HELP PLEASE! (2 replies)
  75. Transformation and ligation not working!!!!!!! (9 replies)
  76. problem in ligation and cloning (1 reply)
  77. Electrophoresis gel boxes and geldoc system (2 replies)
  78. how CIP treatment is done (3 replies)
  79. ampicillin concentration trouble (2 replies)
  80. Fab library constructing-ligation/transformation problem (0 replies)
  81. E. coli strain TB1 vs JM83 (1 reply)
  82. Storage of gel extracted DNA before ligation (3 replies)
  83. Insertion of fragment with same 5' and 3' ends (5 replies)
  84. Dual Luciferase Assay (3 replies)
  85. NICKED DNA (2 replies)
  86. Site directed mutagenesis (1 reply)
  87. cDNA clone (1 reply)
  88. Mini prep (4 replies)
  89. Ligation Problems (2 replies)
  90. problem with pGEMT (3 replies)
  91. Chosing the right competent cells (1 reply)
  92. Checking PCR insert (into pGEMT vector) (2 replies)
  93. Looking for pMSCVneo vector (0 replies)
  94. Qiagen midi prepare low yield (6 replies)
  95. Culture Grow well but No Plasmid (2 replies)
  96. Empty TOPO expression plasmid (4 replies)
  97. Ligation and Transformation help needed: pENTR1A Vector issue (5 replies)
  98. Protein expression with T7 promoter (1 reply)
  99. Checking Plasmid Expression (1 reply)
  100. Triple ligation not working (1 reply)
  101. please urgently help: after phenol/chloroform extraction, my plasmid vector beca (2 replies)
  102. need help cloning cdna of a known protein (1 reply)
  103. Why, oh why, am I having so much trouble with ligations and transformations? (4 replies)
  104. Problem with Induction (inclusion bodies?) (4 replies)
  105. Colony PCR screen positive - insert digestion negative (2 replies)
  106. Which pGL4 vector to choose (2 replies)
  107. plasmid purification kit for transfection (2 replies)
  108. What does the DH5alpha mean in E.coli DH5alpha bacteria (10 replies)
  109. Help: incompatible RE site subcloning (0 replies)
  110. Trouble with subcloning (6 replies)
  111. How does the lac operon in pGEM-T Easy actually work? (0 replies)
  112. restriction problem (3 replies)
  113. pGEMT-easy & blue white screening (4 replies)
  114. overnight restriction digestion- good or bad? (12 replies)
  115. glycerol stocks (10 replies)
  116. please help!!! with transformation (10 replies)
  117. SacI enzyme trouble (2 replies)
  118. same weird weird staph transformation results (12 replies)
  119. Troubleshooting ligation/transformation (1 reply)
  120. TOPO-Transformation Problem (4 replies)
  121. anyone working currently on gateway cloning???? (1 reply)
  122. Promoter Cloning for Luciferase Assay (3 replies)
  123. cloning large insert in pGEM-T easy vector (2 replies)
  124. Lb-kan plates life (2 replies)
  125. trouble with plasmid transformation in BL21DE3 E. coli cells (9 replies)
  126. LR reaction (1 reply)
  127. designing primers for genes not sequenced yet (2 replies)
  128. Cloning Problem (3 replies)
  129. Looking for source for searching Enzyme binding censensus (0 replies)
  130. Help needed for ligation of large inserts into TOPO vector (3 replies)
  131. Can transormations be incubated at less than 37 degrees after heat shock? (4 replies)
  132. AdEasy Vector System: Why PacI digestion? (1 reply)
  133. Transfected protein is constitutively active (0 replies)
  134. Restriction buffer composition (2 replies)
  135. What is the type of replicon from pGC blue plasmid? (2 replies)
  136. Addition of A overhang for dummies? (7 replies)
  137. Yet another ligation problem (4 replies)
  138. Problems with Quick change lightning site directed mutagenesis kit(STRATAGENE) (2 replies)
  139. PCR and then ligation (7 replies)
  140. Signal pIgplus vector sequence (0 replies)
  141. 3 way ligation help needed (10 replies)
  142. cloning protocol (1 reply)
  143. Creating mutants (3 replies)
  144. Adeno-x Tet-on vector problem (0 replies)
  145. Ligation Time and Temp (4 replies)
  146. Miniprep (3 replies)
  147. cloning problems in C. elegans expression plasmid (1 reply)
  148. Mammalian expression vector for a partial protein fragment? (0 replies)
  149. help with cloning using pBudce4.1 (0 replies)
  150. Cloning and Expression Method Survey (1 reply)
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